Microbicidal Pyrimidine or Triazine Containing Compounds

ABSTRACT

The present invention concerns the microbicidal activity of certain pyrimidine or triazine containing non-nucleoside reverse transcriptase inhibitors. The compounds of the present invention inhibit the systemic infection of a human being with HIV, in particular, the present compounds prevent sexual HIV transmission in humans.

The present invention concerns the microbicidal activity of certainpyrimi dine or triazine containing non-nucleoside reverse transcriptaseinhibitors (NNRTIs), in particular, the present invention concerns theuse of pyrimidine or triazine derivatives in the manufacture of amedicament for the prevention of HIV (Human Immunodeficiency Virus)transmission or infection in humans, in particular sexual transmission.It also concerns pharmaceutical compositions adapted to be applied atthe site where sexual intercourse or related intimate contact takesplace.

Worldwide, the heterosexual route is the prevalent mode of transmissionof AIDS. Therefore, demands have been raised for measures that blocksexual spreading of the HIV infection. As there is no effectivetreatment or vaccine against AIDS, preventive measures are the onlytools that can presently reduce transmission of Human Immunodeficiencyvirus (HIV). The consistent and correct use of condoms represents aneffective barrier to prevent HIV transmission. However, the reduction ofacquiring infection can only be significantly reduced if condoms areused for almost all sexual intercourses; a result that can not beachieved despite intensive prevention programs to increase condom use.

Development of microbicides for topical use may represent an efficaciousalternative to condoms. A microbicide is any agent that kills ordeactivates disease-causing microbes. According to the InternationalAssociation of Physicians in AIDS CARE (IAPAC), the definition ofmicrobicides also includes interventions that can block or preventinfection, as well as amplification of the body's natural defenses toprevent infection through sexual acts.

Ideally, microbicides should have little or no side effects at aneffective microbicidal concentration. One aspect in this respect is thatthe drug used as microbicide should have little or no immunosuppressiveactivity at an effective microbicidal concentration. In addition, theideal microbicide should sufficiently withstand varying temperatures andacceptably function within varied pH ranges (ranges of alkaline andacidic levels in the vagina). Further, it should not eliminate thenatural beneficial lactobacilli that reside in the vagina and regulatevaginal health.

Studies have demonstrated that HIV transmission through direct,biological mechanisms are facilitated in a person already infected witha sexually transmitted disease (STD) (Fleming et al. SexuallyTransmitted Infections (1999 February), 75(1), 3-17).

Sores, lesions lesions and inflammations caused by STDs compromisecertain physical barriers to disease, For these reasons, taking measuresto prevent STD transmission is a valuable strategy in the fight againstHIV infection. Several microbicides in human clinical trials containdetergent-type ingredients, which may cause lesions at vaginal andcervical epithelia. Spermicidal products containing biodetergents caninactivate HIV in vitro. However, it has been shown that suchbiodetergents may exacerbate genital ulcers and facilitate HIVtransmission when tested in vivo.

Besides surfactants, which directly act on the virus particle, drugsthat block the early steps of HIV multiplication such as antiretroviraldrugs are undergoing preclinical assessment. Various antiretroviralsincluding non-nucleoside reverse transcriptase inhibitors (NNRTIs) havebeen tested in vitro with varying results. To date, no evidence has beenpublished on the in vivo effectiveness of NNRTIs as microbicidal agents.

It has now been found that the pyrimidine and triazine compounds of thepresent invention exhibit microbicidal activity in that these compoundshave the ability to prevent the infection by HIV.

In addition, the pyrimidine and triazine compounds of the presentinvention also have shown microbicidal activity against STD pathogenssuch as Haeinophilus dicreyi, while maintaining their compatibility withlactobacilli and normal vagina flora. The derived healing effect of thepresent compounds on the chancroids caused by Haemophilus ducreyi,significantly contributes to the prevention of systemic HIV infection.

EP 1002795, WO 99/50250, WO 99/50256 and WO 00/27828 disclose compoundsinhibiting the replication of the HIV virus in the human T-4 cells viaan interaction with the HIV reverse transcriptase enzyme.

DESCRIPTION OF THE INVENTION

The present invention concerns the use of compounds having the formula(I), (II) and (III) wherein a compound of formula (I) corresponds to

a N-oxide, a pharmaceutically acceptable addition salt or astereochernically isomeric form thereof, wherein

Y is CR⁵ or N,

A is CH, CR⁴ or N;

n is 0, 1, 2, 3 or 4;

Q is —NR¹R² or when Y is CR⁵ then Q nay also be hydrogen;

-   -   R¹ and R² are each independently selected from hydrogen,        hydroxy, C₁₋₁₂alkyl, C₁₋₁₂alkyloxy, C₁₋₁₂alkylcarbonyl,        C₁₋₁₂alkyloxycarbonyl, aryl, amino, mono- or        di(C₁₋₁₂alkyl)amino, mono- or di(C₁₋₁₂alkyl)aminocarbonyl        wherein each of the aforementioned C₁₋₁₂alkyl groups may        optionally and each individually be substituted with one or two        substituents each independently selected from hydroxy,        C₁₋₆alkyloxy, hydroxyC₁₋₆alkyloxy, carboxyl,        C₁₋₆alkyloxycarbonyl, cyano, amino, imino, aminocarbonyl,        aminocarbonylamino, mono- or di(C₁₋₆alkyl)amino, aryl and Het;        or    -   R¹ and R² taken together may form pyrrolidinyl, piperidinyl,        morpholinyl, azido or mono- or        di(C₁₋₁₂alkyl)aminoC₁₋₄alkylidene;

R³ is hydrogen, aryl, C₁₋₆alkylcarbonyl, C₁₋₆alkyloxycarbonyl, C₁₋₆alkylsubstituted with C₁₋₆alkyloxycarbonyl; and

-   -   each R⁴ independently is hydroxy, halo, C₁₋₆alkyl, C₁₋₆alkyloxy,        cyano, aminocarbonyl, nitro, amino, trihalomethyl,        trihalomethyloxy, or when Y is CR⁵ then R⁴ may also represent        C₁₋₆alkyl substituted with cyano or aminocarbonyl;    -   R⁵ is hydrogen or C₁₋₄alkyl;    -   L is —X¹—R⁶ or -X²-Alk-R⁷ wherein        -   R⁶ and R⁷ each independently are phenyl or phenyl            substituted with one, two, three, four or five substituents            each independently selected from halo, hydroxy, C₁₋₆alkyl,            C₁₋₆alkyloxy, C₁₋₆alkylcarbonyl , C₁₋₆alkyloxycarbonyl,            formyl, cyano, nitro, amino, and trifluoromethyl; or when Y            is CR⁵ then R⁶ and R⁷ may also be selected from phenyl            substituted with one, two, three, four or five substituents            each independently selected from aminocarbonyl,            trihalomethyloxy and trihalomethyl; or when Y is N then R⁶            and R⁷ may also be selected from indanyl or indolyl, each of            said indanyl or indolyl may be substituted with one, two,            three, four or five substituents each independently selected            from halo, hydroxy, C₁₋₆alkyl, C₁₋₆alkyloxy,            C₁₋₆alkylcarbonyl, C₁₋₆alkyloxycarbonyl, formyl, cyano,            nitro, amino, and trifluoromethyl; when R⁶ is optionally            substituted indanyl or indolyl, it is preferably attached to            the remainder of the molecule via the fused phenyl ring. For            instance, R⁶ is suitably 4-, 5-, 6- or 7-indolyl;        -   X¹ and X² are each independently —NR³—, —NH—NH—, —N═N—, —O—,            —S—, —S(═O)— or —S(═O)₂—;

Alk is C₁₋₄alkanediyl; or

-   -   when Y is CR⁵ then L may also be selected from C₁₋₁₀alkyl,        C₃₋₁₀alkenyl, C₃₋₁₀alkynyl, C₃₋₇cycloalkyl, or C₁₋₁₀alkyl        substituted with one or two substituents independently selected        from C₃₋₇cycloalkyl, indanyl, indolyl and phenyl, wherein said        phenyl, indanyl and indolyl may he substituted with one, two,        three, four or where possible five substituents each        independently selected from halo, hydroxy, C₁₋₆alkyl,        C₁₋₆alkyloxy, cyano, aminocarbonyl, C₁₋₆alkyloxycarbonyl,        formyl, nitro, amino, trihalomethyl, trihalomethyloxy and        C₁₋₆alkylearbonyl;    -   aryl is phenyl or phenyl substituted with one, two, three, four        or five substituents each independently selected from halo,        C₁₋₆alkyl, C₁₋₆alkyloxy, cyano, nitro and triflouromethyl;    -   Het is an aliphatic or aromatic heterocyclic radical; said        aliphatic heterocyclic radical is selected from pyrrolidinyl,        piperidinyl, homopiperidinyl, piperazinyl, morpholinyl,        tetrahydrofuranyl and tetrahydrothienyl wherein each of said        aliphatic heterocyclic radical may optionally be substituted        with an oxo group; and said aromatic heterocyclic radical is        selected from pyrrolyl, furanyl, thienyl, pyridyl, pyrimidinyl,        pyrazinyl and pyridazinyl wherein each of said aromatic        heterocyclic radical may optionally be substituted with hydroxy;    -   and, wherein a compound of formula (II) corresponds to

a N-oxide, a pharmaceutically acceptable addition salt, quaternary amineand the stereochemically isomeric forms thereof, wherein

-b¹=b²-C(R^(2a))=b³-b⁴= represents a bivalent radical of formula

—CH═CH—C(H^(2a))═CH—CH═  (b-1);

—N═CH—C(R^(2a))═CH—CH═  (b-2);

—CH═N—C(R^(2a))═CH—CH═  (b-3);

—N═CH—C(R^(2a))═N—CH═  (b-4);

—N═CH—C(R^(2a))═CH—N═  (b-5);

—CH═N—C(R^(2a))═N—CH═  (b-6);

—N═N—C(R^(2a))═CH—CH═  (b-7);

q is 0, 1, 2; or where possible q is 3 or 4;

-   -   R¹ is hydrogen, aryl, formyl, C₁₋₆alkylcarbonyl, C₁₋₆alkyl,        C₁₋₆alkyloxycarbonyl, C₁₋₆alkyl substituted with formyl,        C₁₋₆alkylcarbonyl, C₁₋₆alkyloxycarbonyl;    -   R^(2a) is cyano, aminocarbonyl, mono- or        di(methyl)aminocarbonyl, C₁₋₆alkyl substituted with cyano,        aminocarbonyl or mono- or di(methyl)aminocarbonyl, C₂₋₆ alkenyl        substituted with cyano, or C₂₋₆alkynyl substituted with cyano;    -   each R² independently is hydroxy, halo, C₁₋₆alkyl optionally        substituted with cyano or —C(═O)R⁶, C₃₋₇cycloalkyl, C₂₋₆alkenyl        optionally substituted with one or more halogen atoms or cyano,        C₂₋₆alkynyl optionally substituted with one or more halogen        atoms or cyano, C₁₋₆alkyloxy, C₁₋₆alkyloxycarbonyl, carboxyl,        cyano, nitro, amino, mono- or di(C₁₋₆alkyl)amino,        polyhalomethyl, polyhalomethyloxy, polyhalomethylthio,        —S(═O)_(p)R⁶, —NH—S(═O)_(p)R⁶, —C(═O)R⁶, —NHC(═O)H, —C(═O)NHNH₂,        —NHC(═O)R⁶, —C(═NH)R⁶ or a radical of formula

-   -   wherein each A independently is N, CH or CR⁶);        -   B is NH, O, S or NR⁶,        -   p is 1 or 2; and        -   R⁶ is methyl, amino, mono- or dimethylamino or            polyhalomethyl;    -   L is C₁₋₁₀alkyl, C₂₋₁₀alkenyl, C₂₋₁₀alkynyl, C₃₋₇cycloalkyl,        whereby each of said aliphatic group may be substituted with one        or two substituents independently selected from        -   C₃₋₇cycloalkyl,        -   indolyl or isoindolyl, each optionally substituted with one,            two, three or four substituents each independently selected            from halo, C₁₋₆allkyl, hydroxy, C₁₋₆alkyloxy, cyano,            aminocarbonyl, nitro, amino, polyhalomethyl,            polyhalomethyloxy and C₁₋₆alkylcarbonyl,        -   phenyl, pyridinyl, pyrimidinyl, pyrazinyl or pyridazinyl,            wherein each of said aromatic rings may optionally be            substituted with one, two, three, four or five substituents            each independently selected from the substituents defined in            R²; or    -   L is —X—R³ wherein        -   R³ is phenyl, pyridinyl, pyrimidinyl, pyrazinyl or            pyridazinyl, wherein each of said aromatic rings may            optionally be substituted with one, two, three, four or five            substituents each independently selected from the            substituents defined in R²; and        -   X is —NR¹—, —NH—NH—, —N═N—, —O—, —C(═O)—, —CHOH—, —S—,            —S(═O)— or —S(═O)₂—;    -   Q represents hydrogen, C₁₋₆alkyl, halo, polyhaloC₁₋₆alkyl or        —NR⁴R⁵; and    -   R⁴ and R⁵ are each independently selected from hydrogen,        hydroxy, C₁₋₆alkyl, C₁₋₁₂alkyloxy, C₁₋₁₂alkylcarbonyl,        C₁₋₁₂alkyloxycarbonyl, aryl, amino, mono- or        di(C₁₋₁₂alkyl)amino, mono- or di(C₁₋₁₂alkyl)aminocarbonyl,        wherein each of the aforementioned C₁₋₁₂alkyl groups may        optionally and each individually be substituted with with one or        two substituents each independently selected from hydroxy,        C₁₋₆alkyloxy, hydroxyC₁₋₆alkyloxy, carboxyl,        C₁₋₆alkyloxycarbonyl, cyano, amino, imino, mono- or        di(C₁₋₆alkyl)amino, polyhalomethyl, polyhalomethyloxy,        polyhalomethylthio, —S(═O)_(p)R⁶, —NH—S(═O)_(p)R⁶, —C(═O)R⁶,        —NHC(═O)H, —C(═O)NHNH₂, —NHC(═O)R⁶, —C(═NH)R⁶, aryl and Het; or    -   R4 and R⁵ taken together may form pyrrolidinyl, piperidinyl,        morpholinyl, azido or mono- or        di(C₁₋₁₂alkyl)aminoC₁₋₄alkylidene;    -   Y represents hydroxy, halo, C₃₋₇cycloalkyl, C₂₋₆alkenyl        optionally substituted with one or more halogen atoms,        C₂₋₆alkynyl optionally substituted with one or more halogen        atoms, C₁₋₆alkyl optionally substituted with cyano or —C(═O)R⁶,        C₁₋₆alkyloxy, C₁₋₆alkyloxycarbonyl, carboxyl, cyano, nitro,        amino, mono- or di(C₁₋₆alkyl)amino, polyhalomethyl,        polyhalomethyloxy, polyhalomethylthio, —S(═O)_(p)R⁶,        —NH—S(═O)_(p)R⁶, —C(═O)R⁶, —NHC(═O)H, —C(═O)NHNH₂, —NHC(═O)R⁶,        —C(═NH)R⁶ or aryl;    -   aryl is phenyl or phenyl substituted with one, two, three, four        or five substituents each independently selected from halo,        C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyloxy, cyano, nitro,        polyhaloC₁₋₆alkyl and polyhaloC₁₋₆alkyloxy;

Het is an aliphatic or aromatic heterocyclic radical; said aliphaticheterocyclic radical is selected from pyrrolidinyl, piperidinyl,homopiperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl andtetrahydrothienyl wherein each of said aliphatic heterocyclic radicalmay optionally be substituted with an oxo group; and said aromaticheterocyclic radical is selected from pyrrolyl, furanyl, thienyl,pyridinyl, pyrimidinyl, pyrazinyl and pyridazinyl wherein each of saidaromatic heterocyclic radical may optionally be substituted withhydroxy; Het is meant to include all the possible isomeric forms of theheterocycles mentioned in the definition of Het, for instance, pyrrolylalso includes 2H-pyrrolyl; the Het radical may be attached to theremainder of the molecule of formula (II) through any ring carbon orheteroatom as appropriate, thus, for example, when the heterocycle ispyridinyl, it may be 2-pyridinyl, 3-pyridinyl or 4-pyridinyl;

and, wherein a compound of formula (III) corresponds to

a N-oxide, a pharmaceutically acceptable addition salt, quaternary amineand the stereochemically isomeric forms thereof, wherein

-a¹=a²-a³=a⁴- represents a bivalent radical of formula

—CH═CH—CH═CH—  (a-1);

—N═CH—CH═CH—  (a-2);

—N═CH—N═CH—  (a-3);

—n═CH—CH═N—  (a-4);

—N═N—CH═CH—  (a-5);

-   -   n is 0, 1, 2, 3 or 4; and in case -a¹=a²-a³=a⁴- is (a-1), then n        may also be 5;    -   R¹ is hydrogen, aryl, formyl, C₁₋₆alkylcarbonyl, C₁₋₆alkyl,        C₁₋₆alkyloxycarbonyl, C₁₋₆alkyl substituted with formyl,        C₁₋₆allkylcarbonyl, C₁₋₆alkyloxycarbonyl, and    -   each R² independently is hydroxy, halo, C₁₋₆alkyl optionally        substituted with cyano or —C(═O)R⁴, C₃₋₇cycloalkyl, C₂₋₆alkenyl        optionally substituted with one or more halogen atoms or cyano,        C₂₋₆alkynyl optionally substituted with one or more halogen        atoms or cyano, C₁₋₆alkyloxy, C₁₋₆alkyloxycarbonyl, carboxyl,        cyano, nitro, amino, mono- or di(C₁₋₆alkyl)amino,        polyhalomethyl, polyhalomethyloxy, polyhalomethylthio,        —S(═O)_(p)R⁴, —NH—S(═O)_(p)R⁴, —C(═O)R⁴, —NHC(═O)H, —C(═O)NHNH₂,        —NHC(═O)R⁴, —C(═NH)R⁴ or a radical of formula

-   -   wherein each A independently is N, CH or CR⁴;        -   B is NH, O, S or NR⁴;        -   p is 1 or 2; and        -   R⁴ is methyl, amino, mono- or dimethylamino or            polyhalomethyl;    -   L is C₁₋₁₀alkyl, C₂₋₁₀alkenyl, C₂₋₁₀alkynyl, C₃₋₇cycloalkyl,        whereby each of said aliphatic group may be substituted with one        or two substituents independently selected from        -   C₃₋₇cycloalkyl,        -   indolyl or isoindolyl, each optionally substituted with one,            two, three or four substituents each independently selected            from halo, C₁₋₆alkyl, hydroxy, C₁₋₆alkyloxy, cyano,            aminocarbonyl, nitro, amino, polyhalomethyl,            polyhalomethyloxy and C₁₋₆allkylcarbonyl,        -   phenyl, pyridinyl, pyrimidinyl, pyrazinyl or pyridazinyl,            wherein each of said aromatic rings may optionally be            substituted with one, two, three, four or five substituents            each independently selected from the substituents defined in            R²; or    -   L is —X—R³ wherein        -   R³ is phenyl, pyridinyl, pyrimidinyl, pyrazinyl or            pyridazinyl, wherein each of said aromatic rings may            optionally be substituted with one, two, three, four or five            substituents each independently selected from the            substituents defined in R²; and        -   X is —NR¹—, —NH—NH—, —N═N—, —O—, —C(═O)—, —CHOH—, —S—,            —S(═O)— or —S(═O)₂—;    -   aryl is is phenyl or phenyl substituted with one, two, three,        four or five substituents each independently selected from halo,        C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyloxy, cyano, nitro,        polyhaloC₁₋₆alkyl and polyhaloC₁₋₆alkyloxy;    -   with the proviso that compounds of formula (III) wherein        -   L is C₁₋₃alkyl; R¹ is selected from hydrogen, ethyl and            methyl; -a¹=a²-a³=a⁴- represents a bivalent radical of            formula (a-1); n is 0 or 1 and R² is selected from fluoro,            chloro, methyl, trifluoromethyl, ethyloxy and nitro; or        -   is —X—R³, X is —NH—; R¹ is hydrogen; -a¹=a²-a³=a⁴-            represents a bivalent radical of formula (a-1); n is 0 or 1            and R² is selected from chloro, methyl, methyloxy, cyano,            amino and nitro and R³ is phenyl, optionally substituted            with one substituent selected from chloro, methyl,            methyloxy, cyano, amino and nitro;    -   and the compounds        -   N,N′-dipyridinyl-(1,3,5)-triazine-2,4-diamine;        -   (4-chloro-phenyl)-(4(1-(4-isobutyl-phenyl)-ethyl)-(1,3,5)            triazin-2-yl)-amine are not included;

in the manufacture of a medicament useful for preventing thetransmission of or infection with HIV, particularly via sexualintercourse or related intimate contact between partners. In particular,the use of a compound of formula (I), (II) or (III) in the manufactureof a topical medicament useful for preventing the transmission of orinfection with HIV.

Thus, the present invention also concerns a method to prevent thetransmission of or infection with HIV, particularly via sexualintercourse or related intimate contact between partners, which methodcomprises administering, in particular topically administering, to ahuman an effective amount, in particular a microbicidal effectiveamount, of a microbici dal compound of formula (I), formula (II) orformula (III).

Suitably, the present invention concerns the use of a compound offormula (I), (II) or (III) in the manufacture of a microbicidalmedicament useful for preventing the transmission of HIV wherein Y inthe compound of formula (II) represents hydroxy, halo, C₃₋₇cycloalkyl,C₂₋₆alkenyl optionally substituted with one or more halogen atoms,C₂₋₆alkynyl optionally substituted with one or more halogen atoms,C₁₋₆alkyl substituted with cyano or —C(═O)R⁶, C₁₋₆alkyloxy,C₁₋₆alkyloxycarbonyl, carboxyl, cyano, nitro, amino, mono- ordi(C₁₋₆alkyl)amino, polyhalomethyl, polyhalomethyloxy,polyhalomethylthio, —S(═O)_(p)R⁶, —NH—S(═O)_(p)R⁶, —C(═O)R⁶, —NHC(═O)H,—C(═O)N H₂, —NHC(═O)R⁶, —C(═NH)R⁶ or aryl.

The term sexual intercourse or related intimate contact between partnerscomprises vaginal sex, anal sex, oral sex and contact of body sites withHIV infected fluids of the sexual partner, in particular semen.Particularly, the term sexual intercourse or related intimate contactbetween partners constitutes vaginal, anal or oral sex, moreparticularly vaginal sex.

The contact sites believed to be most responsible for the transmissionof HIV via sexual intercourse or related intimate contact betweenpartners are the genitals, rectum, mouth, hands, lower abdomen, upperthighs.

The term “partners” as mentioned hereinbefore or hereinafter defines twoor more warm-blooded animals, in particular humans, who are sexuallyactive with each other, ie. who have sexual intercourse with each otheror who have intimate contact with each other related to sexualactivities.

In an embodiment, the present invention concerns the use of compoundshaving the formula (IV), wherein a compound of formula (IV) correspondsto

a N-oxide, a pharmaceutically acceptable addition salt, quaternary amineand the stereochemically isomeric forms thereof, wherein

n is 0, 1, 2, 3 or 4;

-   -   R¹ is hydrogen, aryl, formyl, C₁₋₆alkylcarbonyl, C₁₋₆alkyl,        C₁₋₆alkyloxycarbonyl, C₁₋₆alkyl substituted with formyl,        C₁₋₆alkylcarbonyl, C₁₋₆alkyloxycarbonyl; and    -   R^(2a) is cyano; aminocarbonyl; mono- or dimethylaminocarbonyl;        C₁₋₆alkyl optionally substituted with cyano, aminocarbonyl, or        mono- or dimethylaminocarbonyl; C₂₋₆alkenyl substituted with        cyano; and C₂₋₆alkynyl substituted with cyano;    -   each R² independently is hydroxy, halo, C₁₋₆alkyl optionally        substituted with cyano or —C(═O)R⁴, C₃₋₇cycloalkyl, C₂₋₆alkenyl        optionally substituted with one or more halogen atoms or cyano,        C₂₋₆alkynyl optionally substituted with one or more halogen        atoms or cyano, C₁₋₆alkyloxy, C₁₋₆alkyloxycarbonyl, carboxyl,        cyano, nitro, amino, mono- or di(C₁₋₆alkyl)amino,        polyhalomethyl, polyhalomethyloxy, polyhalomethylthio,        —S(═O)_(p)R⁴, —NH—S(═O)_(p)R⁴, —C(═O)R⁴, —NHC(═O)H, —C(═O)NHNH₂,        —NHC(═O)R⁴, —C(═NH)R⁴ or a radical of formula

-   -   wherein each A independently is N, CH or CR⁴;        -   B is NH, O, S or NR⁴;        -   p is 1 or 2; and        -   R⁴ is methyl, amino, mono- or dimethylamino or            polyhalomethyl;    -   L is C₁₋₁₀alkyl, C₂₋₁₀alkenyl, C₂₋₁₀alkynyl, C₃₋₇cycloalkyl,        each of said aliphatic group substituted with phenyl, which may        optionally be substituted with one, two, three, four or five        substituents each independently selected from the substituents        defined in R²; or    -   L is —X—R³ wherein        -   R³ is phenyl, optionally substituted with one, two, three,            four or five substituents each independently selected from            the substituents defined in R²; and        -   X is —NR¹—, —NH—NH—, —N═N—, —O—, —C(═O)—, —CHOH—, —S—,            —S(═O)— or —S(═O)₂—;    -   aryl is phenyl or phenyl substituted with one, two, three, four        or five substituents each independently selected from halo,        C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyloxy, cyano, nitro,        polyhaloC₁₋₆alkyl and polyhaloC₁₋₆alkyloxy;

with the proviso that the compound 2,4-di-p-cyanoanilino-1,3,5-triazineis not included;

in the manufacture of a microbicidal medicament useful for preventingthe transmission of or infection with HIV.

Thus, the present invention also concerns a method to prevent thetransmission of or infection with HIV, which method comprisesadministering, in particular topically administering, to a human aneffective amount, in particular a microbicidal effective amount, of amicrobicidal compound of formula (IV).

As used in the foregoing definitions and hereinafter halo definesfluoro, chloro, bromo and iodo; polyhalomethyl as a group or part of agroup is defined as mono- or polyhalosubstituted methyl, in particularmethyl with one or more fluoro atoms, for example, difluoromethyl ortrifluoromethyl; polyhaloC₁₋₆alkyl as a group or part of a group isdefined as mono- or polyhalosubstituted C₁₋₆alkyl, for example, thegroups defined in halomethyl, 1,1-difluoro-ethyl and the like; in casemore than one halogen atoms are attached to an alkyl group within thedefinition of polyhalomethyl or polyhaloC₁₋₆alkyl, they may be the sameor different; C₁₋₄alkyl as a group or part of a group encompasses thestraight and branched chained saturated hydrocarbon radicals having from1 to 4 carbon atoms such as, for example, methyl, ethyl, propyl, butyland the like; C₁₋₆alkyl as a group or part of a group encompasses thestraight and branched chained saturated hydrocarbon radicals as definedin C₁₋₄alkyl as well as the higher homologues thereof containing 5 or 6carbon atoms such as, for example pentyl or hexyl; C₁₋₁₀alkyl as a groupor part of a group encompasses the straight and branched chainedsaturated hydrocarbon radicals as defined in C₁₋₆alkyl as well as thehigher homologues thereof containing 7 to 10 carbon atoms such as, forexample, heptyl, octyl, nonyl or decyl; C₁₋₁₂alkyl as a group or part ofa group encompasses the straight and branched chained saturatedhydrocarbon radicals as defined in C₁₋₁₀alkyl as well as the higherhomologues thereof containing 11 or 12 carbon atoms such as, forexample, undecyl, dodecyl and the like; C₁₋₄alkylidene as a group orpart of a group defines bivalent straight and branched chainedhydrocarbons having from 1 to 4 carbon atoms such as, for example,methylene, ethylidene, propylidene, butylidene and the like;C₁₋₄alkanediyl as a group or part of a group encompasses those radicalsdefined under C₁₋₄alkylidene as well as other bivalent straight andbranched chained hydrocarbons having from 1 to 4 carbon atoms such as,for example, 1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl and thelike; C₃₋₇cycloalkyl as a group or part of a group is generic tocyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl;C₃₋₁₀alkenyl as a group or part of a group defines straight and branchchained hydrocarbon radicals containing one double bond and having from3 to 10 carbon atoms such as, for example, 2-propenyl, 2-butenyl,2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl, 3-hexenyl, 3-heptenyl,2-octenyl, 2-nonenyl, 2-decenyl and the like, whereby the carbon atomattached to the pyrimidine ring is preferably an aliphatic carbon atom;C₃₋₁₀alkynyl as a group or part of a group defines straight and branchchained hydrocarbon radicals containing one triple bond and having from3 to10 carbon atoms such as, for example, 2-propynyl, 2-butynyl,2-pentynyl, 3-pentynyl, 3-methyl-2-butynyl, 3-hexynyl, 3-heptynyl,2-octenyl, 2-nonynyl, 2-decenyl and the like, whereby the carbon atomattached to the pyrimidine ring is preferably an aliphatic carbon atom;C₂₋₆alkenyl defines straight and branched chain hydrocarbon radicalshaving from 2 to 6 carbon atoms containing a double bond such asethenyl, propenyl, butenyl, pentenyl, hexenyl and the like; C₂₋₁₀alkenyldefines straight and branched chain hydrocarbon radicals having from 2to 10 carbon atoms containing a double bond such as the groups definedfor C₂₋₆alkenyl and heptenyl, octenyl, nonenyl, decenyl and the like;C₂₋₆alkynyl defines straight and branched chain hydrocarbon radicalshaving from 2 to 6 carbon atoms containing a triple bond such asethynyl, propynyl, butynyl, pentynyl, hexynyl and the like; C₂₋₁₀alkynyldefines straight and branched chain hydrocarbon radicals having from 2to 10 carbon atoms containing a triple bond such as the groups definedfor C₂₋₆alkynyl and heptynyl, octynyl, nonynyl, decynyl and the like;C₁₋₃alkyl as a group or part of a group encompasses the straight andbranched chain saturated hydrocarbon radicals having from 1 to 3 carbonatoms such as, methyl, ethyl and propyl; C₄₋₁₀alkyl encompasses thestraight and branched chain saturated hydrocarbon radicals as definedabove, having from 4 to 10 carbon atoms. The term C₁₋₆a defines straightor branched chain saturated hydrocarbon radicals such as methoxy,ethoxy, propyloxy, butyloxy, pentyloxy, hexyloxy, 1-methylethyloxy,2-methylpropyloxy, 2-methylbutyloxy and the like; C₃₋₆cycloalkyloxy isgeneric to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy andcyclohexyloxy.

As used herein before, the term (═O) forms a carbonyl moiety whenattached to a carbon atom, a sulfoxide group when attached once to asulfur atom, and a sulfonyl group when attached twice to a sulfur atom.

When any variable (e.g. aryl etc.) occurs more than one time in anyconstituent, each definition is independent.

Lines drawn into ring systems from substituents indicate that the bondmay be attached to any of the suitable ring atoms. For instance forcompounds of formula (I), R⁴ can be attached to any available carbonatom of the phenyl or pyridyl ring.

For use in the presently described medicaments and methods, salts of thecompounds of the present invention are those wherein the counted on ispharmaceutically acceptable. However, salts of acids and bases which arenon-pharmaceutically acceptable may also find use, for example, in thepreparation or purification of a pharmaceutically acceptable compound.All salts, whether pharmaceutically acceptable or not are includedwithin the ambit of the present invention.

The pharmaceutically acceptable addition salts as mentioned hereinaboveare meant to comprise the microbicidal active non-toxic addition saltforms which the compounds of the present invention are able to form. Thelatter can conveniently be obtained by treating the base form with suchappropriate acids as inorganic acids, for example, hydrohalic acids,e.g. hydrochloric or hydrobromic acid and the like; sulfuric acid;nitric acid; phosphoric acid and the like; or organic acids, forexample, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic,malonic, succinic, maleic, fumaric, malic, tartaric, citric,methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluene-sulfonic,cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.Conversely the salt form can be converted by treatment with alkali intothe free base form.

The pharmaceutically acceptable addition salts as mentioned hereinaboveare also meant to comprise the microbicidal active non-toxic base forms,in particular, metal or amine addition salt forms which the compounds ofthe present invention are able to form. Said salts can conveniently beobtained by treating the compounds of the present invention containingacidic hydrogen atoms with appropriate organic and inorganic bases suchas, for example, the ammonium salts, the alkali and earth alkaline metalsalts, e.g. the lithium, sodium, potassium, magnesium, calcium salts andthe like, salts with organic bases, e.g. primary, secondary and tertiaryaliphatic and aromatic amines such as methylamine, ethylamine,propylamine, isopropylamine, the four butylamine isomers, dimethylamine,diethylamine, diethanolamine, dipropylamine, diisopropylamine,di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine,triethylamine, tripropylamine, quinuclidine, pyridine, quinoline andisoquinoline, the benzathine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, hydrabamine salts, and saltswith amino acids such as, for example, arginine, lysine and the like.Conversely said salt forms can be converted by treatment with acid intothe free acid form.

The term addition salts comprises as well the hydrates and the solventaddition forms which the compounds of the present invention are able toform. Examples of such forms are e.g. hydrates, alcoholates and thelike.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds are able to form by reaction betweena basic nitrogen of a compound and an appropriate quaternizing, agent,such as, for example, an optionally substituted alkylhalide, arylhalideor arylalkylhalide, e.g. methyliodide or benzyliodide. Other reactantswith good leaving groups may also be used, such as alkyltrifluoromethanesulfonates, alkyl methanesulfonates, and alkylp-toluenesulfonates. A quaternary amine has a positively chargednitrogen. Pharmaceutically acceptable counterions include chloro, bromo,iodo, trifluoroacetate and acetate. The counterion of choice can beintroduced using ion exchange resins.

The N-oxide forms of the present compounds are meant to comprise thecompounds wherein one or several tertiary nitrogen atoms are oxidized tothe so-called N-oxide.

The term stereochemically isomeric forms of the compounds of the presentinvention, their N-oxides, addition salts, quaternary amines, as usedhereinbefore, defines all possible compounds made up of the same atomsbonded by the same sequence of bonds but having differentthree-dimensional structures which are not interchangeable, which thecompounds of the present invention may possess. Unless otherwisementioned or indicated, the chemical designation of a compoundencompasses the mixture of all possible stereochemically isomeric formsthat said compound may possess. Said mixture may contain alldiastereomers and/or enantiomers of the basic molecular structure ofsaid compound. All stereochemically isomeric forms of the compounds bothin pure form or in admixture with each other are intended to be embracedwithin the scope of the present invention.

In particular, stereogenic centers may have the R- or S-configuration;substituents on bivalent cyclic (partially) saturated radicals may haveeither the cis- or trans-configuration. Compounds encompassing doublebonds can have an E (entgegen) or Z (zusammen)-stereochemistry at saiddouble bond. The terms cis, trans, R, S, E and Z are well known to aperson skilled in the art.

Some of the present compounds may also exist in their tautomeric forms.Such forms although not explicitly indicated in the above formula areintended to be included within the scope of the present invention.

Whenever used hereinafter, the term “compounds”, the term “compounds ofthe present invention” is meant to include any subgroup thereof, alsothe N-oxide forms, the pharmaceutically acceptable addition salts, thequaternary amines and all stereochemically isomeric forms. Of specialinterest are those compounds which are stereochemically pure.

Whenever substituents can be selected each independently from a list ofnumerous definitions, such as for example for R⁶ and R⁷, all possiblecombinations are intended which are chemically possible and which leadto chemically stable molecules.

Suitable compounds of formula (I) are those wherein Y is CR⁵ or N; A isCH, CR⁴ or N; n is 0, 1, 2, 3 or 4; Q is —NR¹R²; R¹ and R² are eachindependently selected from hydrogen, hydroxy, C₁₋₁₂alkyl,C₁₋₁₂allkyloxy, C₁₋₁₂alkylcarbonyl, C₁₋₁₂allkyloxycarbonyl, aryl, amino,mono- or di(C₁₋₁₂alkyl)amino, mono- or di(C₁₋₁₂alkyl)aminocarbonylwherein each of the aforementioned C₁₋₁₂alkyl groups may optionally andeach individually be substituted with one or two substituents eachindependently selected from hydroxy, C₁₋₆alkyloxy, hydroxyC₁₋₆alkyloxy,carboxyl, C₁₋₆alkyloxycarbonyl, cyano, amino, amino, aminocarbonyl,aminocarbonylamino, mono- or di(C₁₋₆alkyl)amino, aryl and Het; or R¹ andR² taken together may form pyrrolidinyl, piperidinyl, morpholinyl, azidoor mono- or di(C-₁₋₁₂alkyl)aminoC₁₋₄alkylidene; R³ is hydrogen, aryl,C₁₋₆alkylcarbonyl, C₁₋₆alkyl, C₁₋₆alkyloxycarbonyl, C₁₋₆alkylsubstituted with C₁₋₆alkyloxycarbonyl; each R⁴ independently is hydroxy,halo, C₁₋₆alkyl, C₁₋₆alkyloxy, cyano, aminocarbonyl, nitro, amino,trihalomethyl, trihalomethyloxy; R⁵ is hydrogen or C₁₋₄alkyl; L is—X¹—R⁶ or -X²-Alk-R⁷ wherein R⁶ and R⁷ each independently are phenyl orphenyl substituted with one, two, three, four or five substituents eachindependently selected from halo, hydroxy, C₁₋₆alkyl, C₁₋₆alkyloxy,C₁₋₆alkylcarbonyl, C₁₋₆alkyloxycarbonyl, formyl, cyano, nitro, amino,and trifluoromethyl, X¹ and X² are each independently —NR³—, —NH—NH—,—N═N—, —O—, —S—, —S(═O)— or —S(═O)₂—, and Alk is C₁₋₄alkanediyl; aryl isphenyl or phenyl substituted with one, two, three, four or fivesubstituents each independently selected from halo, C₁₋₆alkyl,C₁₋₆alkyloxy, cyano, nitro and triflouromethyl; Het is an aliphatic oraromatic heterocyclic radical; said aliphatic heterocyclic radical isselected from pyrrolidinyl, piperidinyl, hornopiperidinyl, piperazinyl,morpholinyl, tetrahydrofuranyl and tetrahydrothienyl wherein each ofsaid aliphatic heterocyclic radical may optionally be substituted withan oxo group; and said aromatic heterocyclic radical is selected frompyrrolyl, furanyl, thienyl, pyridyl, pyrimidinyl, pyrazinyl andpyridazinyl wherein each of said aromatic heterocyclic radical mayoptionally be substituted with hydroxy.

Most preferred compounds of formula (I) are

4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]benzonitrile;

6-[(2,6-dichlorophenyl)methyl]-N2-(4-fluoropheny)-2,4-pyrimidinediamine;

4-[[4-[(2,4-dichlorophenyl)methyl]-6-[(4-hydroxybutyl)amino]-2-pyrimidinyl]amino]-benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(3-hydroxypropyl)amino]-2-pyrimidinyl]-amino]benzonitrile;

N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]-acetamide,

N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]-butanamide;

4-[[2-amino-6-(2,6-dichlorophenoxy)-4-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2-hydroxy-2-phenylethyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[3-(2-oxo-1-pyrrolidinyl)propyl]amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(2-hydroxyethoxy)ethyl]amino]-2-pyrimidinyl]amino]berizontrilemonohydrochloride;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2,3-dihydroxypropyl)amino]-2-pyrimidinyl]-amino]benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-(hydroxyamino)-2-pyrimidinyl]amino]-benzonitrile;

4-[[4-[(2-cyanoethyl)amino]-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]-benzonitrile;

4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(1-pyrroldinyl)ethyl]amino]-2-pyrimidinyl]-amino]benzonitrile;

4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;

N2-(4-bromophenyl)-6-[(2,6-dichlorophenyl)methyl]-5-methyl-2,4-pyrimidinediamine;

4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[2-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-(2,4,6-trimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dichlorophenyl)thio]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[[2,6-dichloro-4-(trifluoromethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,4-dichloro-6-methylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[2-[(cyanophenyl)amino]-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;

4-[[4-[(2,4-dibromo-6-fluorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-5-methyl-2-pyrimidinyl]amino]-benzeneacetonitrile;

4-[[4-[methyl(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,4,6-trichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,4,6-trimethylphenyl)amino-2-pyrimidinyl]amino]benzonitrile;

4-[[4-amino-6-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[2-amino-6-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;

4-[[4-(2-bromo-4-chloro-6-methylphenoxy)-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(4-chloro-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

3,5-dichloro-4-[[2-[(4-cyanophenyl)amino]-4-pyrimidinyl]amino]benzonitrile;

4-[[4-[[2,6-dichloro-4-(trifluoromethoxy)phenyl]amino]-2-pyrimidinyl]amino]-benzonitrile;

4-[[4-[(2,4-dibromo-3,6-dichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dibromo-4-propylphenyl]amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzamide;

4-[[4-[(4-(1,1-dimethylethyl)-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[2-[(4-cyanophenyl)amino]4-pyrimidinyl]oxy]-3,5-dimethylbenzonitrile;

4-[[4-[(4-chloro-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;

4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino-3,5-dimethylbenzonitrile;

4-[[4-[[4-(1,1-dimethylethyl)-2,6-dimethylphenyl]amino]-5-methyl-2-pyrimidinyl]-amino]benzonitrile;

4-[[4-[(4-bromo-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;

4-[[5-methyl-4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-[(2,6-dibromo-4-propylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;

4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzamide,N3-oxide;

N2-(4-chlorophenyl)-N4-(2,4,6-trimethylphenyl)-2,4-pyrimidinediamine;

4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;

4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;

4-[[4-[(phenylmethyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-amino-6-(2,6-dimethylphenoxy)-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-[(2-chloro-6-methylphenyl)amino]-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-[(2,4,6-trimethylphenyl)amino]-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-(hydroxyamino)-6-[(2,4,6-trimethylphenyl)amino]-1,3,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-amino-6-[(2-ethyl-6-methylphenyl)amino]-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-[(2,6-dichlorophenyl)thio]-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-(hydroxyamino)-6-[(2,4,6-trichlorophenyl)amino]-1,3,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-amino-6-(2,4,6-trimethylphenoxy)-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-(hydroxyamino)-6-(2,4,6-trimethylphenoxy)-1,3,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-amino-6-[(2,4-dichloro-6-methylphenyl)amino]-13,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-[(2,4-dichloro-6-methylphenyl)amino]-6-(hydroxyamino)-1,3,5-triazin-2-yl]-amino]benzonitrile;

4-[[4-(hydroxyamino)-6-(2,4,6-trichlorophenoxy)-1,3,5-triazin-2-yl]amino]benzonitriletrifluoroacetate (1:1);

4-[[4(4-acetyl-2,6-dimethylphenoxy)-6-amino-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-(2,4,6-tribromophenoxy)-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-(4-nitro-2,6-dimethylphenoxy)-1,3,5-tiazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-(2,6-dibromo-4-methylphenoxy)-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-(4-formyl-2,6-dimethylphenoxy)-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-amino-6-[(2,4-dichlorophenyl)thio]-1,3,5-triazin-2-yl]amino]benzonitrile;

4-[[4-[(5-acetyl-2,3-dihydro-7-methyl-1H-inden-4-yl)oxy]-6-amino-1,3,5-triazin-2-yl]-amino]benzonitrile;

4-[[4-amino-6-[(4-bromo-2-chloro-6-methylphenyl)amino]-1,3,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-amino-6-[(2-chloro-4,6-dimethylphenyl)amino]-1,3,5-triazin-2-yl]amino]-benzonitrile;

4-[[4-amino-6-[[2,4-dichloro-6-(trifluoromethyl)phenyl]amino]-1,3,5-triazin-2-yl]-amino]benzonitrile;

4-[[4-amino-6-[methyl(2,4,6-trimethylphenyl)amino]-1,3,5-triazin-2-yl]-amino]-benzonitrile;

4-[[4-amino-6-[(2,6-dibromo-4-methylphenyl)amino]-1,3,5-triazin-2-yl]-amino]-benzonitrile;

4-[[4-amino-6-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-1,3,5-triazin-2-yl]-amino]benzonitrile,

the N-oxides, the pharmaceutically acceptable addition salts and thestereochemically isomeric forms thereof.

Suitable compounds of forint/la (II) are those wherein one or more ofthe following restrictions apply:

-   -   -b¹=b²-C(R^(2a))=b³-b ⁴= is a radical of formula (b-1);    -   q is 0;    -   R^(2a) is cyano or —C(═O)NH₂, preferably R^(2a) is cyano;    -   Y is cyano, —C(═O)NH₂ or a halogen, preferably a halogen;    -   Q is hydrogen or —NR⁴R⁵ wherein R⁴ and R⁵ are preferably        hydrogen;    -   L is —X—R³ wherein X is preferably —NR¹—, —O— or —S—, most        preferably X is —NH—, and R³ is substituted phenyl with        C₁₋₆alkyl, halogen and cyano as preferred substituents.

Another interesting group of compounds of formula (II) are thosecompounds wherein L is —X—R³ wherein R³ is 2,4,6-trisubstituted phenyl,each substituent independently selected from chloro, bromo, fluoro,cyano or C₁₋₄alkyl.

Also interesting are those compounds of formula (II) wherein Y is chloroor bromo and Q is hydrogen or amino.

Particular compounds are those compounds of formula (II) wherein themoiety in the 2 position of the pyrimidine ring is a 4-cyano-anilinogroup.

Preferred compounds are those compounds of formula (II) wherein themoiety in the 2 position of the pyrimidine ring is a 4-cyano-anilinogroup, L is —X—R³ wherein R³ is a 2,4,6-trisubstituted phenyl, Y is ahalogen and Q is hydrogen or NH₂,

Most preferred compounds of formula (II) are:

4-[[4-amino-5-chloro-6-[(2,4,6-trimethylpheny)amino]-2-pyrimidinyl]amino]-benzonitrile;

4-[[5-chloro-4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[5-bromo-4-(4-cyano-2,6-dimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;

4-[[4-amino-5-chloro-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;

4-[[5-bromo-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;

4-[[4-amino-5-chloro-6-(4-cyano-26-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile;and

4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile;the N-oxides, the pharmaceutically acceptable addition salts, quaternaryamities and the stereochemically isomeric forms thereof.

An interesting group of compounds are those compounds of formula (III)wherein one or more of the following conditions are met:

-   -   n is 1;    -   -a¹=a²-a³=a⁴- represents a bivalent radical of formula (a-1);    -   R¹ is hydrogen or C₁₋₆alkyl;    -   R² is cyano; aminocarbonyl; mono- or di(methyl)aminocarbonyl;        C₁₋₆alkyl substituted with cyano, aminocarbonyl or mono- or        di(methyl)aminocarbonyl; and more in particular, R² is on the 4        position relative to the —NR¹— moiety;    -   L is —X—R³ wherein X is preferably —NR¹—, —O— or —S—, most        preferably X is —NH—, and R³ is substituted phenyl with        C₁₋₆alkyl, halogen and cyano as preferred substituents.

Preferred compounds are those compounds of formula (III) wherein L is—X—R³ wherein R³ is a disubstituted phenyl group or a trisubstitutedphenyl group, each substituent independently selected from chloro,bromo, fluoro, cyano or C₁₋₄alkyl.

Most preferred compound of formula (III) is4-[[4-[(2,4,6-trimethylphenyl)amino]-1,3,5-triazin-2-yl]amino]benzonitrile.

Particular compounds compounds of the present invention include4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]benzonitrile(compound A) and4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile(compound B), their N-oxides, pharmaceutically acceptable salts andstereoisomers thereof.

The compounds of the present invention can be prepared according toart-known procedures. In particular, they are prepared according to theprocedures described in EP 1002795, WO 99/50250, WO 99/50256 and WO00/27828.

The compounds of the present invention have microbici dal activity andhave the ability to prevent the transmission of HIV. In particular, theycan prevent sexual or vaginal transmission of HIV by preventing eitherthe production of infectious viral particles or infection of uninfectedcells. If infected cells in sperm can reach the mucosa, the compounds ofthe present invention can prevent HIV infection of host cells, such asmacrophages, lymphocytes, Langerhans and M cells. Thus, the presentcompounds prevent systemic HIV infection of a human being, exhibiting aprophylactic action against HIV. Evidence for this microbicidal activityis given in the experimental part and is based on in vivo activity ofcompound B in a human SCID (Severe Combined Immune Deficiency) animalmodel (Di Fabio et al., AIDS 2001, 15, 2231-2238) and on in vitroactivity of Compound B in a model based on immature monocyte deriveddendritic cells.

In addition, it has been found that the compounds of this invention havea killing effect on the Haemophilus ducreyi bacteria. As such, thecompounds of this invention may be used in the prevention and treatmentof chancroids, the venereal disease caused by this bacteria. Theseadditional effects will even improve the effectiveness of the presentcompounds in preventing infection with HIV.

The compounds of the invention may be formulated into pharmaceuticalcompositions that can be used to apply microbicides to effectivelyprevent transmission of pathogens through mucosae and/or skin, moreparticularly to prevent the sexual or vaginal transmission of HIV. Thus,the compositions are in forms adapted to be applied to the site wheresexual intercourse or related intimate contact takes place, such as thegenitals, vagina, vulva, cervix, rectum, mouth, hands, lower abdomen,upper thighs, especially the vagina, vulva, cervix, and ano-rectalmucosae.

The compounds of the present invention may be formulated intopharmaceutical compositions designed for immediate release or sustainedor slow release.

As appropriate topical compositions there may be cited for example gels,jellies, creams, pastes, emulsions, dispersions, ointments, films,sponges, foams, aerosols, powders, intravaginal rings or otherintravaginal drug delivery systems, cervical caps, implants, patches,suppositories or pessaries for rectal, or vaginal application, vaginalor rectal or buccal tablets, mouthwashes.

To prepare the pharmaceutical compositions of this invention, aneffective amount of the particular compound, optionally in addition saltform, as the active ingredient may be combined in intimate admixturewith a pharmaceutically acceptable carrier, which carrier may take awide variety of forms depending on the form of administration. Forexample, in preparing the compositions for topical oral administration,any of the usual pharmaceutical media may be employed such as, forexample, water, glycols, oils, alcohols and the like, which are suitablefor oral liquid preparations such as mouthwashes in the form ofsuspensions, emulsions and solutions. Solid carriers such as starches,sugars, kaolin, diluents, lubricants, binders, disintegrating agents andthe like will be adequate in the case of tablets. Also included aresolid form preparations which are intended to be converted, shortlybefore use, to liquid form preparations. In the compositions suitablefor topical cutaneous administration, the carrier optionally comprises asuitable wetting agent, optionally combined with suitable additives ofany nature in minor proportions, which additives do not introduce asignificant deleterious effect on the skin. Said additives mayfacilitate the administration to the skin and/or may be helpful forpreparing the desired compositions. These compositions may beadministered in various ways, e.g., as a cream or gel.

The active ingredient may be present in the pharmaceutical formulationsas a free agent or alternatively, encapsulated into drug carriers likeliposomes, nanoparticles or cyclodextrins, which encapsulation resultsin an increased concentration of the compounds within the microbe targetsite. The active ingredient may also be present as nanoparticles.

Liposomes may be present in the formulation which include, amongstothers, distearoylphosphatidylcholine (DSPC),distearoylphosphatidyglycerol (DSPG),distearoylphosphatidylethanolamine-polyethylene-glycol (DSPE-PEG),dipalmitoylphosphatidylcholine (DPPC), dicetylphosphate (DP),cholesterol (CHOL), dipalmitoylphosphatidylglycerol (DPPG), andcombinations thereof, such as distearoylphosphatidylcholine (DSPC):distearoylphosphatidylglycerol (DSPC); within which the activeingredient is entrapped.

Appropriate cyclodextrins are α-, β-, γ-cyclodextrins or ethers andmixed ethers thereof wherein one or more of the hydroxy groups of theanhydroglucose units of the cyclodextrin are substituted with C₁₋₆alkyl,particularly methyl, ethyl or isopropyl, e.g. randomly methylated β-CD;hydroxyC₁₋₆alkyl, particularly hydroxyethyl, hydroxypropyl orhydroxybutyl; carboxy C₁₋₆alkyl, particularly carboxymethyl orcarboxyethyl; C₁₋₆alkylcarbonyl, particularly acetyl. Especiallynoteworthy as complexants and/or solubilizers are β-CD, randomlymethylated β-CD, 2,6-dimethyl-β-CD, 2-hydroxyethyl-β-CD,2-hydroxyethyl-β-CD, 2-hydroxypropyl-β-CD and(2-carboxymethoxy)propyl-β-CD, and in particular 2-hydroxypropyl-62 -CD(2-HP-β-CD). Cyclodextrins are additionally useful in enhancing thesolubility of the compounds.

The term mixed ether denotes cyclodextrin derivatives wherein at leasttwo cyclodextrin hydroxy groups are etherified with different groupssuch as, for example, hydroxy-propyl and hydroxyethyl.

Particularly, the present compounds may be formulated as a gelformulation comprising:

-   -   a topically effective amount of a compound of the present        invention;    -   a gel-forming compound;    -   a buffer;    -   a pharmaceutically acceptable diluent, preferably water;    -   optionally a humectant; and    -   optionally a preservative.

Typical gel formulations can be prepared using natural or syntheticpolymers as gelifying agents, and hydrophobic or hydrophilic liquids.Examples of gel-forming compounds commonly employed in gel formulationsinclude polysaccharides which include cellulose derivatives,glycosaminoglycans, gums, starch (a-amylose or amylopectin), andchitosan; carboxyvinylic derivates, vinyl polymers such aspolyethylenes, polyehtyelene glycols, e.g. polyethylene glycol 4500,Plastibase® (Plasticized Hydrocarbon Gel), polyacrylic acid, (Carbopols®family, e.g. Carbopol® 940), polymethacrylic acid, polyvinyl pyrrolidoneand polyvinyl alcohol; polyacrylamide or polymethacrylamide polymersincluding clays such as bentonite, Veegum® (R.T Vanderbilt) andLaponite® (Laporte Industries); polyoxyethylene-polyoxypropylene orpolyethylene oxides copolymers such as poloxamers, e.g. poloxamer 407,poloxamines; proteins, colloidal silica, soaps, silicones such asdimethylpolysiloxanes or dimeticone, hydrocarbonated bases (mixtures ofparafine and yaselines).

Useful cellulose derivates include methyl cellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose,carboxymethyl cellulose. Useful glycosaminoglycans include hyaluronicacid, chondroitin, chondroitin-4-sulfate, heparan sulfate and heparin.Useful gums include natural and artificial gums, tragacanth,carrageenan, pectin, agar, alginic acid, dextrans. Theglycosaminoglycans may be used to enhance wound healing in combinationwith any other gel-forming polymer such as, for example, collagen,gelatin, fibronectin. A preferred gelifying agent is hydroxyethylcellulose, which has additionally bioadhesive properties.

Concentrations of the gel-forming compounds may be varied uponconditions such as the liquid/gel transition temperature, the physicalproperties sought for the gel and the pH used in the making of theformulations.

Gel forming compounds employed in the present invention are typicallywater-soluble polymers capable of forming a viscous aqueous solution, ornon-water soluble, water-swellable polymers (e. g., collagen) that canalso form a viscous solution and that gel upon contact with skin.Gelling agents suitable for use in the present invention should bestable over a wide pH range, especially over the normal acidic pH valuesfound in the vagina.

Buffering agents are used in the gel formulation of this invention tomaintain the pH of the vagina within its healthy acidic range (i.e., apH of less than about 5 and more preferably in the range of about 3.2 toabout 4.5) even in the presence of normal amounts of ejaculate. A normalacidic range in the vaginal milieu and environment assists indiminishing the activity of certain STD-causing microbes, including theHIV virus. Maintaining the normal vaginal milieu also assists inmaintaining the body's natural defenses against certain STD-causingmicroorganisms. Examples of buffering agents include, without beinglimited to, lactic acid, phosphoric acid, sodium citrate, sodiumhydroxide, sodium phosphate, sodium phosphate dibasic anhydrous,tartaric acid, triethanolamine, citric acid, potassium acid tartrate,benzoic acid, alginic acid, sorbic acid, fumaric acid, ascorbic acid,stearic acid, oleic acid, edetic acid, ethylenediaminetetracetic acid,acetic acid, malic acid, and the like, preferably sodium hydroxide andlactic acid, the latter being additionally a preservative and havingcertain antimicrobial activity.

The acids acids may be added as free acids, hydrates, orpharmaceutically acceptable salts. Free acids can be converted to thecorresponding salts in situ (i.e., within the vagina). It is generallypreferred that several buffering agents are included in the gel of thisinvention to provide increased buffering capacity. Even more preferably,buffering agents comprise a combination of acid and hydrogen-acceptingsubstance that occur naturally in the human female body that, whenapplied to the surface of the vagina, maintains the pH level thereon atapproximately the pH level of a healthy vagina. Said acid(s) may beselected from the froup consisting of acetic acid, lactic acid,phosphoric acid and sulfuric acid, and combinations thereof. One of thecharacteristics common to each of said member in said group, is thateach acid occurs naturally in the female body. Another commoncharacteristic is each readily contributes to the formation of abuffering system, by temporarily donating hydrogen ion and accepting acation to form a salt.

Said hydrogen-accepting substance may be selected from the groupconsisting of potassium hydroxide, sodium hydroxide, calcium hydroxide,potassium carbonate, sodium carbonate and calcium carbonate, andcombinations thereof. One of the characteristics common to each of saidmember in said group, is that each substances is found naturally in thefemale body. Another common characteristic is each readily contributesto the formation of a buffering system, by temporarily acceptinghydrogen ion and donating a cation to form a salt. Such salts may beselected from the group consisting of acetate, lactate, phosphate andsulfate, in combination with said cation from said hydrogen-acceptingsubstance.

The gels of this invention may also include, and preferably do include,humectants. Suitable humectants include, for example, glycerol,polyethylene glycols, propylene glycols, sorbitol, triacetin, and thelike. Glycerol, which is the preferred humectant, is a buffer activatingcomponent due to its capability of water absortion, or other fluid, fromthe vaginal environment into the gel. It is believed that such fluidintake prevents the formation of a dry film on the gel when placedwithin the vagina, providing additional solvent to enhance theapplication of the gel formulation, or to otherwise enhance itsfunctioning.

The gels of this invention may also include, and preferably do include,a preservative, which amongst other properties, extends the shelf lifeof the gel formulations. Suitable preservatives include, for example,benzoic acid, sodium benzoate, methyl paraben, ethyl paraben, butylparaben, propyl paraben, benzylalkonium chloride, phenylmercuricnitrate, chlorhexidine, benzyl alcohol, phenethyl alcohol, propyleneglycol, and the like. The preferred preservatives are methyl paraben,and propyl paraben, which both also contribute to the antimicrobialcapacity of the gel.

The gels of this invention are prepared using conventional gelpreparation techniques. It is desirable, however, to ensure that thebuffering agents are solubilized in the final product and that theentrapment of air in the gel is avoided or at least kept to a minimum.To reduce the entrapment of air in the gel, it is generally preferredthat the less hydrophilic agents are added in small increments.Alternatively, the gels of this invention can also be prepared inreadily dispersable solid forms (e.g., powders, tablets, and the like)which can be converted to the desired gel consistency by action ofaqueous based fluids external to or within the vagina when desired. Asthose skilled in the art will realize, the methods for preparing thegels of this invention can be modified for batch, semi continuous, orcontinuous operation so long as the resulting gels have the desired andbeneficial properties described herein.

The gel formulations can be combined with other active ingredients suchas microbicides, antimicrobials, chemotherapeutic agents,antiinflammatory agents, spermicides or other appropriate drugs.Furthermore, microbicides or spermicides or both can be combined withliposomes (or other drug carriers) to prevent any disease of mucosaeand/or skin. In addition, gel or liposome or other drug carriersformulations can also be used as carriers of vaccines against infectionscaused by pathogens or any disease. If desired, flavorants, scents,fragrances, and colorants can be incorporated into the gel so long asthey do not interfere with the protection afforded by the gel. Indeed,incorporation of such flavorants, scents, fragrances, and colorants intothe compositions of this invention may provide further protection byincreasing the probability that the gel will be used during sexualactivity.

In one embodiment, the gel formulation is composed of compound B,hydroxyethyl cellulose (HEC), glycerol, methyl paraben, propyl paraben,lactic acid, sodium hydroxide (for reaching a pH around 4.5), and water.

In another embodiment, the gel formulation comprises compound B, HECwith a concentration from about 0.5 to about 5% (w/w), glycerol with aconcentration from about 1 to about 15% (w/w), methyl paraben with aconcentration from about 0.02 to about 0.5% (w/w), propyl paraben with aconcentration from about 0.005 to about 0.2% (w/w), lactic acid with aconcentration from about 0.005 to about 0.5% (w/w), sodium hydroxide insufficient quantity to achieve a pH of 4.5, and water.

In another embodiment, the gel formulation comprises compound B, HECwith a concentration from about 1 to about 3% (w/w), glycerol with aconcentration from about 3 to about 7% (w/w), methyl paraben with aconcentration from about 0.1 to about 0.3% (w/w), propyl paraben with aconcentration from about 0.01 to about 0.03% (w/w), lactic acid with aconcentration from about 0.03 to about 0.07% (w/w), sodium hydroxide insufficient quantity to achieve a pH of 4.5, and water.

In another embodiment, any of the above gel formulations comprisecompound A as a microbicide.

The present topical formulations such as the gel formulations describedherein are to be used for coating different types of mucosae such asvulvar, vaginal, cervical, ano-rectal, mouth, or skin to prevent thepenetration of pathogens such as viruses, bacteria, fungi, parasites,ectoparasites and mycoplasmas.

The present topical formulations such as the gel formulations describedherein could, for example, be applied into the vagina by hand,suppositories, or conventional tampon or syringe techniques. The methodof administering or delivering the gel into the vagina is not criticalso long as an effective amount of the gel is delivered into the vagina.The present topical formulations such as the gel formulations describedherein may also be used for protection during anal intercourse and canbe applied using similar techniques.

For vaginal heterosexual intercourse, the present topical formulationssuch as the gel formulations described herein may be applied into thevagina prior to intercourse. For anal intercourse (heterosexual orhomosexual), the present topical formulations such as the gelformulations described herein may be inserted into the rectum prior tointercourse. For either vaginal or anal intercourse, the present topicalformulations such as the gel formulations described herein may also actas a lubricant. For added protection it is generally preferred that thepresent topical formulations such as the gel formulations describedherein be applied before intercourse or other sexual activity and that,if appropriate, a condom be used. For even further protection, thepresent topical formulations such as the gel formulations describedherein can be applied as soon as possible after completion of the sexualactivity. Although application only after the sexual activity is lessrecommended, it would still be desirable afterwards if the applicationwas not performed prior to the sexual activity for any reason (e.g., incases of rape).

The present topical formulations such as the gel formulations describedherein are highly suited for the protection of women (as well as theirpartners) with or without requiring the partner's knowledge of theapplication of these gels. In addition, reliance on the partner's claimof being STD-free, concretely HIV-free, would not be necessary, neitherthe agreement to use condoms or other barrier devices for protection.

The gel formulations of the present invention are additionallyadvantageous because they do not significantly affect or inhibit thegrowth characteristics of the normal vaginal flora or otherwisesignificantly irritate the vaginal tissue when used at inhibitory,noncytotoxic, or clinical concentrations. Significant inhibition ormodifications of the vaginal flora or other irritations can lead toincreased risks of infections (both STD and non-STD types) frequentlymediated by ulcerations in the vagina, unusual discharges, generaldiscomforts, and the like.

Intravaginal rings (IVR) are as well suitable drug delivery systems forthe vaginal administration of the compounds of the present invention.IVRs comprise the compound(s) dispersed throughout a biocompatibleelastomeric system that forms the delivery device, which preferentiallytakes the form of a ring. These elastomers preferably includehydrophobic material, such as silicones (organo polysiloxanes includingdimethylpolysiloxanes), polyethylene-co-poly (vinyl acetate),styrene-butadiene-styrene block copolymers, polyphosphazenes,poly(isoprene), poly (isobutylene), polybutadienes, polyurethanes,nitrile rubbers, neoprene rubbers or mixtures thereof. Said IVRs can beformulated as sustained-released microbicides, resulting in an extendedand stable contact time between the compound and target pathogens andcells. IVR formulations have already been described in the literature,WO02076426 all of which is herein incorporated by reference.

In order to increase the residence time of the topical pharmaceuticalcomposition at the site of administration, it may be advantageous toinclude a bioadhesive in the different drug delivery systems, inparticular a bioadhesive polymer. A bioadhesive may be defined as amaterial that adheres to a living biological surface such as for examplea mucus membrane or skin tissue. The term bioadhesive is well-known tothe person skilled in the art. Thus, the present invention also relatesto a pharmaceutical composition comprising a pharmaceutically acceptablecarrier and as active ingredient a microbicidal effective amount of acompound of the invention characterized in that the pharmaceuticalcomposition is bioadhesive to the site of application. Preferably, thesite of application is the vagina, vulva, cervix, rectum, mouth or skin,most preferred is the vagina and the vulva.

Examples of bioadhesives which may be used in the pharmaceuticalcompositions of the present invention comprise polyacrylic acidderivatives, such as for example carbopol or polycarbophil, e.g.carbopol 934P, carbopol 940, polycarbophil AA1; cellulose etherderivatives such as for example hydroxypropyl methylcellulose,hydroxypropyl cellulose, hydroxyethyl cellulose, sodium carboxymethylcellulose, chitosan; natural polymers such as for example alginates,tragacanth, inulin; pregelatinized starch; polysaccharide gums such asxanthan gum, and the like.

Alternatively, formulations of the present invention may be in the formof implants, patches, pads, injections or other preparations forachieving a percutaneous and subcutaneous delivery of the compounds tothe cervical, vaginal and rectal tissues.

As already indicated within the gel specifications, the presentcompounds may be used in all the suitable formulations, alone or incombination with other active ingredients, such as antivirals,antibiotics, immunomodulators or vaccines. They may also be used aloneor in combination with other prophylactic agents for the prevention ofviral infections. The present compounds may be used in vaccines andmethods for protecting individuals against viral infections over anextended period of time. The compounds may be employed in such vaccineseither alone or together with other compounds of this invention ortogether with other anti-viral agents in a manner consistent with theconventional utilization of reverse transcriptase inhibitors invaccines. Thus, the present compounds may be combined withpharmaceutically acceptable adjuvants conventionally employed invaccines and administered in prophylactically effective amounts toprotect individuals over an extended period of time against HIVinfection.

Antiviral compounds which may be used in combination with the compoundsof the invention may be known antiretroviral compounds such as suramine,pentamidine, thymopentin, castanospermine, dextran (dextran sulfate),foscarnet-sodium (trisodium phosphono formate); nucleoside reversetranscriptase inhibitors, e.g. zidovudine (3′-azido-3′-deoxythymidine,AZT), didanosine (2′,3′-dideoxyinosine; ddI), zalcitabine(dideoxycytidine, ddC) or lamivudine (2′-3′-dideoxy-3′-thiacytidine,3TC), stavudine (2′,3′)-didehydro-3′-deoxythymidine, d4T), abacavir andthe like; non-nucleoside reverse transcriptase inhibitors such asnevirapine(11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido-[3,2-b:2′,3-e][1,4]diazepin-6-one), efavirenz, delavirdine, and the like; phosphonatereverse transcriptase inhibitors, e.g. tenofovir and the like; compoundsof the TIBO (tetrahydro-imidazo[4,5,1-jk][1,4]-benzodiazepine-2(1H)-oneand thione)-type(S)-8-chloro-4,5,6,7-tetrahydro-5-methyl-6-(3-methyl-2-butenyl)imidazo-[4,5,1-jk][1,4]benzo-diazepine-2(1H)-thione,compounds of the α-APA (α-anilino phenyl acetamide) type e.g.α-[(2-nitrophenyl)amino]-2,6-dichlorobenzene-acetamide and the like;inhibitors of trans-activating proteins, such as TAT-inhibitors, e.g.RO-5-3335, or REV inhibitors, and the like; protease inhibitors e,gindinavir, ritonavir, saquinavir, lopinavir (ABT-378), nelfinavir,amprenavir, TMC-126, BMS-232632, VX-175 and the like; fusion inhibitors,e.g. T-20, T-1249 and the like; CXCR4 receptor antagonists, e.g.AMD-3100 and the like; inhibitors of the viral integrase; ribonucleotidereductase inhibitors, e.g. hydroxyurea and the like.

Combinations may as well exert a synergistic effect in inhibiting HIVreplication when components of the combination act on different or samesites of HIV replication, preferably on different sites. The use of suchcombinations may reduce the dosage of a given conventionalantiretroviral agent which would be required for a desired prophylacticeffect as compared to when that agent is administered as a single activeingredient. These combinations reduce potential of resistance to singleagent, while minimizing any associated toxicity. These combinations mayalso increase the efficacy of the conventional agent without increasingthe associated toxicity.

Thus, the compounds of the present invention may also be administered incombination with art-known microbicides, consequently potentiating theprophylactic effect. They can block the infection by creating a barrierbetween the pathogen, in this case the Human Immunodeficiency Virus, andthe site at which transmission will take place, e.g. vulva, vagina; theycan kill or immobilize the pathogen; they can prevent a virus fromreplicating once it has infected the cells lining the site oftransmission, e.g. the cells that line the vaginal wall. Examples ofmicrobicides are:

-   -   Antibiotic peptides: small protein molecules that form part of        the body's first line of defense against infection. These        peptides line every surface of the body—eyes, skin, lungs,        tongue and intestinal tract—and kill bacteria within minutes of        contact. Thus, if applied at the site of potential infection of        HIV, peptides may kill pathogens before they cause infection.    -   Antibodies: isolated antibodies that counteract HIV are        available in the literature, They may be appropriately combined        with the compounds of the present invention to prevent HIV        infection.    -   pH regulators, especially for the vagina. A natural vaginal        environment is too acidic for HIV to survive, but semen        decreases its acidity, allowing HIV to survive. pH regulators        regulate the natural acidity of the vagina making it        inhospitable for the HIV, Said regulators encompass the use of        Lactobacillus bacteria that produce hydrogen peroxide and        thereby help to keep the vaginal environment healthy and acidic,        The acidic polymer BufferGel (ReProtect, LLC) is another example        of a pH regulator which has in addition spermicidal activity.    -   Detergents and surfactants: these compounds are able to disrupt        the outer shell of viruses and therefore are useful as        microbicide and they can be combined with the compounds to        prevent HIV infection. Examples of such detergents and        surfactants are nonoxynol-9 and octoxynol-9, but all detergents        and surfactants that are commonly used in shampoos, toothpastes        and cleaning solutions, contact lens solutions may be equally        suitable.    -   Coatings for the pathogen, such as Pro-2000 Gel which contains a        synthetic polymer that binds to HIV, disrupting the binding of        the virus to target cells.    -   Coatings for the site of transmission, such as for example gels.        These products may prevent HIV from entering the cells by        covering the site of transmission, e.g. the vaginal and vulvar        epithelium. Examples, including the gel preparations described        above, encompass sulphated and sulphonated polymers such as        PC-515 (carrageenan), dextrin 2 sulphate, secretory leukocyte        protease inhibitor (SLPI), which binds to the target cells so        that they are not accessible to the virus, cyanovirin-N which        also binds to the cell, prohibiting cell fusion with HIV.

In the compositions of the present invention, one or more or all of theabove-listed microbicides may be combined with a compound of theinvention. Thus, the present invention also relates to a pharmaceuticalcomposition comprising a compound of the present invention and furthercomprising one or more components wherein the components are selectedfrom antibiotic peptides, antibodies, pH regulators, detergents orsurfactants, coatings for the pathogen, coatings for the site ofadministration.

One particular example of the combination of microbicides is thecombination of compounds of the invention with cellulose acetatephthalate (CAP) and/or hydroxypropyl methylcellulose phthalate (HPMCP).CAP and its derivates are excipients which exhibit an additionalmicrobicide effect, CAP formulations have already been described in theliterature, EP1030547, U.S. Pat. No. 6,165,493, by Neurath et al., allof which are herein incorporated by reference.

The present invention relates also to a pharmaceutical composition asoutlined hereinabove further comprising a spermicidal compound. Saidcompositions are able to prevent at the same time conception and HIVinfection. Suitable spermicides are for example nonoxynol-9,octoxynol-9, menfegol, benzalkoniuni chloride, N-docasanol.

Those of skill in the prophylaxis of HIV-infection could determine themicrobicidal effective amount from the test results presented here andmay range from about 1 ng to about 10 mg, in particular from about 10 ngto about 1 mg, more in particular from about 100 ng to about 100 μg andpreferably from about 500 ng to about 50 μg of active ingredient perapplication or unit dose, in particular an application or unit dose ofan immediate release formulation.

It may be appropriate to apply the required dose as unit dosage forms.The volume of a unit dose, in particular the unit dose of an immediaterelease formulation, whether or not in a unit dosage form, may range inthe case of a topical formulation from about 10 μl up to about 25 ml oftopical formulation and in particular from about 1 ml up to about 10 mlof topical formulation. In the case of a gel or a cream for instance, aconvenient unit dose could range between about 1 ml and about 5 ml.

For instance in the case of topical formulations, in particular topicalformulations for immediate release, as mentioned herein, e.g. a gel, acream and the like, the active ingredient may be present in aconcentration ranging from about 1 nM up to about 10 mM, in particularfrom about 10 nM up to about 1 mM, more in particular from about 100 nMup to about 100 μM and preferably from about 1 μM to about 100 μM.

It is evident that said effective amount may be lowered or increaseddepending on the particular compound being used, on the response of thetreated subject and/or depending on the evaluation of the physicianprescribing the compounds of the instant invention.

EXAMPLES

The following examples are intended to illustrate the present invention.

Example 1: In Vitro Evaluation of the Non-Nucleoside ReverseTranscriptase Inhibitors Compound B as HIV Microbicide

Cell-Free and Cell-Associated HIV Strains

For experiments with CEM T cells, we used the lymphotropic SI/X4 HIVstrain HTLV-III_(B), originally obtained from R. C. Gallo and M. Popovic(NIH, Bethesda, Md.). For experiments with monocyte-derived dendriticcells (MO-DC), the monotropic NSI/R5 HIV strain Ba-L, kindly provided bythe NIH AIDS Research and Reference Reagent Program (Rockville, Md) wasused. Ba-L stocks were grown and tittered on PHA/IL-2-stimulatedperipheral blood mononuclear cells (PBMC) in complete medium, containingRPMI-1640 (Bio-Whittaker, Verviers, Belgium) and 10% bovine calf serum(Hyclone, Utah, US) (Peden K. Virological and Molecular GeneticTechniques for Studies of Established HIV Isolates. 1995, 21-45). Thesupernatant of these cultures were directly used as cell-free virus toinfect MO-DC. To prepare cell-associated HIV Ba-L, non-stimulated PBMC(2×10⁶ cells/ml) were incubated overnight with 10⁻² MOI (=multiplicityof infection) of HIV Ba-L in complete medium. Afterwards, cells wereextensively washed, frozen in liquid nitrogen and thawed on the day ofinfection.

HIV Antigen Detection After Primary Culture and Calculation of EC50Value

HIV antigen was detected using an in-house developed ELISA assay, ofwhich the characteristics have been described (Beirnaert E, Willems B,Peeters M, Bouckaert A, Heyndrickx L, Zhong P et al. Design andEvaluation of an in-House HIV-1 (Group M and O), SIVmnd and SIVcpzAntigen Capture Assay. J Virol Methods 1998, 73: 65-70). The lowerdetection limit is about 200 pg/ml and the upper limit is about 25,000pg/ml, as determined using a standard curve of Ba-L stock dilutions withknown p24 content. The 50% Effective Concentration (EC50) was calculatedby plotting HIV Ag concentration against drug concentration, followed byregression analysis on the linear part of the curve,

Measuring 50% Effective and 50% Ctotoxic Concentrations in CEM T Cells

As a reference system, CEM T cells (obtained from the American Typeculture Collection in Rockville, Md.) were used under previouslystandardized conditions (Balzarini J, et al. AIDS Res Hum Retroviruses10 Apr. 2000, 16: 517-528). Briefly, cells were suspended at 250,000cells/ml in RPMI-1640, supplemented with 10% fetal calf serum, 2 mML-glutamine and 0.075% NaHCO₃ and infected with HTLV-III_(B) at ˜20TCID₅₀. 100 μl of a 5-fold dilution series of the drugs were immediatelyadded to 100 μl of the infected cells in 200-μl well plates. After 4 to5 days of incubation at 37° C., the cultures were examined for syncytiumformation. The EC₅₀ is the concentration required to inhibit syncytiumformation by 50%. Cytotoxicity was evaluated and given as CC50 values,which is the concentration at which the viability of the CEM cells isdecreased by 50%.

Generation of Monocvte-Derived Interstitial-Type Dendritic Cells (MO-DC)and T Cells

Monocytes and lymphocytes were separated from buffy coat PBMC bycounter-flow elutriation, as previously described (Van Herrewege et al.AIDS Res Hum Retroviruses 10 Oct. 2002, 18: 1091-1102). Monocytes werefurther differentiated to MO-DC by culture at 37° C. and 5% CO2 during 7days in complete medium, supplemented with 20 ng/ml GM-CSF and IL-4(Lmmunosource, Zoersel, Belgium) (Sallusto et al. J Exp Med 1 Apr. 1994,179: 1109-1118. Romani et al. J Exp Med 1 Jul. 1994, 180: 83-93.Geissmann F, et al. Exp Med 16 Mar. 1998, 187: 961-966). The lymphocytefraction was frozen in liquid nitrogen and thawed on the day ofinfection. CD4(+) T cells were purified by positive selection, using aCD4(+) isolation kit (Dynal, Oslo, Norway), as described (Vanham et al.AIDS 20 Oct. 2000, 14: 2299-231 1. Vanham et al. AIDS 18 Aug. 2000, 14:1874-1876).

Pre-treatment of HIV with Drugs, with or without Continued Treatment ofMO-DC/CD4(+) T Cell Co-Cultures After Infection

Cell-free HIV Ba-L was pre-incubated with a serial dilution of drug,ranging from 10,000 to 0.1 nM (final concentration), for 1 hour at 37°C. MO-DC were infected with drug-treated HIV at a multiplicity ofinfection (MOI) of 10⁻³. After 2 hours (at 37° C.), MO-DC were washed(6×) and suspended at 4×10⁵ cells/ml. 50 μl of MO-DC were dispensed in a96-well cup, together with 50 μl of autologous CD4(+) T cells (2×10⁶cells/ml) and 100 μl of complete medium or 100 μl of drug (at the sameconcentration as for the pre-incubation). Half of the culture medium wasrefreshed twice weekly with complete medium, with or without drug. After2 weeks of primary culture, supernatants were analysed by ELISA andcells were used for secondary cultures to check viral rescue. Forexperiments with cell-associated HIV, a similar set-up was used exceptthat pre-incubated, cell-associated virus was washed before addition ofMO-DC/CD4(+) T cells and remained present during the MO-DC/CD4(+) T cellco-culture.

24 Hours Drug Treatment of MO-DC/CD4(+) T Cell Co-Cultures During HIVInfection

To evaluate the effect of a 24-hours treatment, MO-DC and autologousCD4(+) T cells were suspended in complete medium at 4×10⁴, resp. 2×10⁵cells/ml. Fifty μl of MO-DC and 50 μl of CD4(±) T cells were dispensedin a 96-well cup, together with 50 μl of cell-associated or cell-freevirus (10⁻³ MOI) and 50 μl of complete medium or 50 μl of a serialdilution of drug. After 24-hours (37° C., 5% CO₂), cells were washed(3×) and incubated for 2 weeks. Half of the culture medium was refreshedtwice weekly with complete medium (without drug). After 2 weeks ofprimary culture, supernatants were analysed by ELISA and cells were usedfor secondary cultures to check viral rescue.

Detection of Viral Rescue: Secondary Culture and PCR Analysis

PBMC were isolated from donor buffy coats and cultured for 2 days incomplete medium supplemented with 5 ng/ml IL-2 (Immunosource, Zoersel,Belgium) and 0.5 μg/ml PHA (Murex, Dartford, England). After 2 weeks ofprimary culture, MO-DC/CD4(+) T cell co-cultures were washed (3×) andsecondary cultures were set-up by adding 1×10⁵ PHA/IL-2 activated PBMCper cup. Half of the culture medium was replaced every 3-4 days withIL-2 containing medium (without drug) and supernatants as well as cellswere harvested after 2 additional weeks. Supernatants were tested forHIV-Ag in ELISA. Cells were processed for HIV DNA measurement, using aPCR-based HIV proviral DNA quantitation kit, developed from AmplicorHIV-1 Monitor™ Test, version 1.5 (Roche Molecular Systems, Branchburg,USA), the modifications of which have been described (Christopherson etal. J Clin Microbiol 2000, 38: 630-634). A lower threshold of 10 HIVcopies per 10⁶ cells was confirmed by using 8E5/LAV cells, containing 1copy of proviral DNA per cell (kindly provided by the CentralizedFacility for AIDS Reagents, Potters Bar, UK)

Evaluation of the Immune Suppressive Activity and Cellular Toxicity ofCompound B in MO-DC/CD4(+) T Cell Co-Cultures

The immune suppressive activity of compound B was measured in mixedleukocyte cultures (MLC), with MO-DC as stimulators and allogenic CD4(+)T cells as responders. Cultures of 3×10³ MO-DC and 100×10³ T cells wereset-up in 6-fold in a 96-well plate, in the presence or absence of adilution series of compound B. In a first part of experiments, compoundB was removed after 24 hours by washing and cells were cultured for anadditional 4 days. In a second part of experiments, compound B remainedpresent during the 5-day culture period. In both set-ups, 1 μCi of[methyl-³H] thymidine (TRA. 120 from Amersham Pharmacia,Buckinghamshire, U.K.) was added to each well at the fifth day ofculture. Plates were harvested 7 hours later and [methyl-³H] thymidineincorporation was measured in a scintillation counter (Top Count™,Canberra-Packard, Zellik, Belgium) and expressed as counts per minute(CPM). The

Immune Suppressive Concentration (ISC₅₀) is defined as the drugconcentration inhibiting 50% of the lymphocyte proliferation. Cellulartoxicity was evaluated microscopically by cosine staining of co-culturesof MO-DC and allogeneic CD4(+) T cells , cultured for 5 days in thepresence of a dilution series of drug. Part of the harvested cells wasalso used for flowcytometric analysis of lymphocyte blast formation andapopoptosis, based on forward and side scatter.

Reference Data on Antiviral Activity and Cellular Toxicity of Compound B

The CEM T cell line was used as a reference to determine the antiviralactivity of compound B. As shown in Table 1, compound B was active inthe nanomolar range and showed a low toxicity. Next to antiviralactivity in CEM T cells, inhibition of HIV-1 reverse transriptaseactivity was measured in a cell-free assay, in which the 50% inhibitoryconcentration (IC50) of said compound is indicated (FIG. 1). The CEMsystem, using a lab T cell-line and the SI/X4labstrain HTLV-IIIb, wasnot directly relevant to sexual transmission, where primary T cells,dendritic cells and NSI/R5 viruses are involved. Therefore, we focussedon prevention of NSI/R5 HIV infection in MO-DC/CD4(+) T cellco-cultures.

TABLE 1 Antiviral activity, cytotoxicity and HIV-1 RT inhibitorycapacity of compound B EC50 CC50 IC50 Drug Treatment HIV (nM) ^(a) (nM)^(b) (nM) ^(c) compound B Continuous HTLV- 1 1.367 24 IIIb ^(a) EC50:50% Effective Concentration, concentration required to inhibit syncytiumformation of HTLV-IIIb infected CEM T cells by 50% ^(b) CC50: 50%Cytotoxic Concentration, concentration at which the viability of CEM Tcells is decreased by 50% ^(c) IC50: 50% Inhibitory Concentration,concentration that inhibits HIV-1 reverse transcriptase activity by 50%.

Drug Treatment of MO-DC/CD4(+) T Cell Co-Cultures Prevented HIVIntegration

In preliminary experiments the HIV Ba-L virus was pretreated for 1 hourwith up to 10,000 nM of compound B. The drug remained present during the2 hours incubation of the virus with the MO-DC, but it was thoroughlywashed away before addition of autologous CD4(+) T cells.

In order to study the maximal effect of the drug, pre-treatment of thevirus and treatment of the cells during infection was combined withfurther treatment during the entire primary culture period of 2 weeks.An example of the inhibitory effects of compound B on infection withcell-associated virus is shown in Table 2. Compound B blocked infectionin the primary cultures already at 10 nM, but addition of PHA/IL-2blasts revealed that 100 nM was needed to completely block infection andprevent proviral integration. When cell-free virus was used forinfection, continuous treatment with 10 nM of compound B sufficed tocompletely block HIV infection, also during secondary culture (Table 3).

Next, it was investigated whether drug treatment during the first 24hours of the primary culture could suffice to prevent viral infectionand integration, as measured by ELISA and PCR respectively. As comparedto continuous treatment, compound B showed to block infection at 1 loghigher concentrations as used for the continuous treatment (Table 3).

TABLE 2 Inhibition of infection of MO-DC/CD4(+) T Cell co-cultures withcell-associated HIV Ba-L HIV Antigen (number of positive wells)^(c) HIVproviral DNA ^(d) Drug Conc (nM) 1° Cult.^(a) 2° Cult.^(b) (2° Cult.)compound B 10,000 0 0 Neg 1,000 0 0 Neg 100 0 0 Neg 10 0 3 4.74 No drug0 6 6 4.85 ^(a)Cell-associated HIV Ba-L was pre-incubated with drug,washed, and added to co-cultures of MO-DC and autologous CD4+ T cells.Cells were cultured for 2 weeks, in the continuous presence of drug(Primary (1°) Culture) ^(b)After primary culture, cells were washed andPHA/IL-2 activated PBMCs were added and maintained in IL-2 containingmedium during a secondary (2°) culture of 2 weeks (no drug present).^(c)Culture supernatant was tested for HIV antigen by ELISA. The numberof antigen-positive microcultures (out of 6) is represented. ^(d) Aftersecondary culture, cells were analysed in PCR for the presence ofproviral DNA, results are expressed as Log(number of DNA copies/10⁶cells)

TABLE 3 Conditions for Prevention of HIV infection in MO-DC/CD4(+) TCell co-cultures Drug Treatment HIV Conc. (nM) ^(a) compound B 24 HoursCell-free 100 Cell-associated 1,000 Continuous Cell-free 10Cell-associated 100 ^(a) MO-DC/CD4(+) T cell co-cultures were incubatedwith cell-free or cell-associated HIV and concurrently drug treatedduring 24 hours or continuously during 1° culture. After 1° culture,cells were washed and used for 2° cultures (no drug present). Drugconcentrations that prevent replicative HIV infection, as measured byELISA of culture supernatants and PCR of cells after 2° culture, areshown. ^(b) The concentration of 10,000 nM was not used in this part ofthe experiment

Compound B had a Low Cellular Toxicity in CEM T Cells and a FavourableTherapeutic Index in MO-DC/CD4(+) T Cell Co-Cultures

Cellular toxicity (CC50 value) in reference GEM T cells was at 1,367 nMfor compound B (Table 1).

The immune suppressive activity of compound B was evaluated in mixedleukocyte cultures with N40-DC as stimulators and allogenic CD4(+) Tcells as responders. If drug was present during the whole cultureperiod, the 50% Immune Suppressive Concentration (ISC50) was about 1,500nM. If the drug was only present during the first 24 hours, the ISC50increased to almost 25,000 nM (Table 4). Thus, the immune suppressiveactivity of compound B was clearly less suppressive in the 24 hourstreatment as compared to continuous treatment. In order to fullyevaluate the relation of anti-viral and immune suppressive activity, the50% antiviral activity (or EC50) values were calculated on primarydrug-treated cultures of HIV-infected autologous MO-DC/CD4(+) T cellco-cultures and the therapeutic indices (TI) were determined. The dataof Table 4 shows that compound B has a favourable TI as measured in thismodel of primary target cells of sexual transmission.

TABLE 4 Overview of the Antiviral and Immune Suppressive Activity ofcompound B in co-cultures of MO-DC/CD4(+) T cells EC50 ISC50 DrugTreatment HIV (nM) ^(a) (nM) ^(b) TI ^(c) Comp. B 24 hours Cell-free42   24,886 592 Cell-associated 63   395 Continuous Cell-free <0.11,515 >15,150 Cell-associated <1 ^(d) >1,515 ^(a) EC50: 50% EffectiveConcentration: drug concentration inhibiting 50% of HIV Ba-L replication^(b) ISC50: 50% Immune Suppressive Concentration: drug concentrationinhibiting 50% of T-lymphocyte proliferation. ^(c) TI: TherapeuticIndex: ISC50/EC50

Additional experiments were done to evaluate if inhibition of DNAsynthesis corresponded to increased mortality of T cells or only todecreased blast formation. Fifty percent inhibition of blast formationwas observed at 3,916 nM. The 50% death rate of T cells was calculatedat 54,222 nM.

In conclusion, prevention of HIV infection was possible against bothcell-free and cell-associated NSI/R5 virus and a 24 hours treatment wassufficient. Compound B showed a high therapeutic index, based on itsrelative weak immune suppressive and potent anti-viral activity. Theseresults confirmed the use of compound B as microbicide.

Example 2: Human SCID Animal Model

In order to mimic the in vivo transmission that occurs in humans, ahu-SCID animal model of vaginal transmission of HIV for the evaluationof vaginal microbicides was used (Di Fabio et al., AIDS 2001, 15,2231-2234 Gels made up of carbopol 940 or hydroxyethyl cellulose (HEC),two water soluble polymers, were prepared containing Compound B indifferent concentrations (0.225 mM; 0.0225 mM or 0.00225 mM). Animalsreceived a single intravaginal application of 25 μl of gel containingCompound B, 15-20 minutes prior to a non invasive vaginal challenge with2×10⁶ human peripheral blood lymphocytes PBL, (hu-PBL) previouslyinfected in vitro with non-syncitium (NSI) strains of HIV-1 (SF162 and1/BX08). Cell to cell transmission was assessed by p24 production and byquantitative PCR. As a result of this study with Compound B, systemicinfection was successfully inhibited.

Example 3: In Vitro Model Based on T-Cell Derived Jurkat-Tat Cells(PM-1)

Direct antiviral activity was demonstrated in a model using Jurkat-tatcells. HIV-1^(RF) (10³TCID₅₀) immobilised into 96-well coated plateswere treated with test compound B for 1 hour at 37° C., compound wassubsequently removed by washing with 4 volumes of PBS before co-culturewith indicator cells for 8 days. Protection of infection was provided atthe 100 nM concentration. Additionally, protection of infection wasdemonstrated at 10 nM in a parallel setup were virus was pretreatedprior to addition of cells and without removal of the compound.

Example 4: Cervical Explant Model

Compound B, at a concentration of 10 nM, was able to block infection ofthe tissue and at a concentration of 100 nM the compound could preventtransfer of infectious virus from migratory dendritic cells toco-cultured T-cells.

Compound demonstrated good efficacy against primary HIV strains (X4,CCR5 and X41R5) in relevant cell lines for a vaginal/rectal microbicideindication: In cervical epithelial cells (MEI180) exposed to thecompound during either 1 hr, 24 hrs or 5 days after which drug wasremoved by washing, viability of the cells was assessed with an MTTassay. Data showed no toxicity at the 50 μM concentration and somereduction of viability at 100 nM.

Example 5: Biochemical Characterization of the Interaction BetweenCompound B and HIV-1 Reverse Transcriptase

In order to investigate the nature of the interaction between compound Band HIV-1 reverse transcriptase (HIV-1 RT), the inhibition of theRNA-dependent DNA polymerization reaction was investigated under steadystate conditions. In a first experiment, the reaction velocity wasdetermined in the presence of different concentrations of compound B anddifferent concentrations of dGTP while the concentration of thep(rC)p(dG) complex was constant. The result showed that the binding ofcompound B to HIV-1 RT is non-competitive against dGTP with a Km valueof 2.51 μM and a Ki of 0.033 μM.

In a second experimental the reaction velocity was determined in thepresence of different concentrations of compound B and differentconcentrations of p(rC)p(dG) while the concentration of the substratewas constant. The result showed that the binding of compound B to HIV-1RT is also non-competitive against p(rC)p(dG) with a Km value of 10.3 μMand a Ki of 0.028 μM. Taken together this means that the binding ofcompound B on HIV-1 RT is independent from the binding of nucleotide andindependent from the binding of primer/template.

Example 6: Compatibility with Lactobacilli and Normal Vaginal Flora

Compatibility with lactobacilli and normal vaginal flora is an importantrequirement for a vaginal microbicide. Activity on pathogens of sexualtransmitted diseases is an additional advantage. In in vitro tests forantibacterial activity of compound B, the following susceptibilitieswere found:

-   -   33 different Lactobacillus species isolated from recto-vaginal        cultures of pregnant women were studied. Results showed a        minimum inhibitory concentration (MIC₅₀) of >32 mg/L    -   Hemophilus ducreyi was inhibited by the compound with a MIC₅₀ of        1 mg/L and a MIC₅₀ of 2 mg/L    -   Some Neisseria gonorrhoea strains were inhibited at clinical        relevant concentrations.

Example 7: Rabbit Vaginal Irritation Test

Several formulations of the compound were used to exclude any irritationin a Rabbit Vaginal irritation test. 24 hours after the application of 1ml of gellcream at different concentrations (0.1M, 0.9 mM, 0.45 mM and0.225 mM), vaginal epithelium was carefully examined macroscopically andmicroscopically. Microscopic slides of parafinized samples of differentparts of the cervico-vagina were stained and analysed histologically.Macroscopic and microscopic scores obtained indicated that theformulations tested were well tolerated.

Example 8: White Rabbit Vaginal Irritation and Toxicity Study

Gel formulations of the compound B at different concentrations were usedto study the exclusion of any irritation in a Rabbit Vaingal Irritationtest. 10 days after the application of the gel formulations onto 6different rabbit groups, vaginal and cervical epithelium were carefullyexamined by a pathologist macroscopically and histologically. Thefollowing scores were determined.

TABLE 5 Vaginal examination Epithelial loss and atrophy accompanied byminimal or slight epithelial inflammatory cell infiltration was onlyseen in all female rabbits treated with Nonoxynol-9, 4%. Group/Sex 1F 2F3F 4F 5F 6F Treatment Sham Placebo gel gel gel Nonoxynol Control(HEC-gel) 22.5 μM 225 μM 10 mM 9, 4% Epithelial loss Marked 0 0 0 0 0 2and atrophy Severe 0 0 0 0 0 3 Total 0 0 0 0 0  5b Epithelial Minimal 00 0 0 0 3 inflammatory Slight 0 0 0 0 0 2 infiltrate Total 0 0 0 0 0  5aap < 0.05 (Fisher's exact two-tailed probability test) bp < 0.01(Fisher's exact two-tailed probability test)

TABLE 6 Cervical examination Loss and atrophy of the epithelia andluminal inflammatory cells and cellular debris were only seen in femalestreated with Nonoxynol-9, 4%. Group/Sex 1F 2F 3F 4F 5F 6F Treatment ShamPlacebo gel gel gel Nonoxynol Control (HEC-gel) 22.5 μM 225 μM 10 mM 9,4% Epithelial loss Slight 0 0 0 0 0 1 and atrophy Marked 0 0 0 0 0 3Total 0 0 0 0 0  4a Luminal Present 0 0 0 0 0 4 Inflammatory Total 0 0 00 0  4a cells ap < 0.05 (Fisher's exact two-tailed probability test)

Example 9: Microbicide Gels

This example illustrates various gels with different dosages of activeprinciple that can be prepared and used as microbicides in theprevention of HIV infection for topical administration.

TABLE 7 22.5 μM gel compound B 0.74 mg HEC 2 g Glycerol 5 g Methylparaben 180 mg Propyl paraben 20 mg Lactic acid 50 mg Sodium hydroxideq.s. for pH 4.5 Water q.s. 100 g

TABLE 8 225 μM gel compound B 7.40 mg HEC 2 g Glycerol 5 g Methylparaben 180 mg Propyl paraben 20 mg Lactic acid 50 mg Sodium hydroxideq.s. for pH 4.5 Water q.s. 100 g

TABLE 9 1 mM gel Compound B 32.94 mg HEC 2 g Glycerol 5 g Methyl paraben180 mg Propyl paraben 20 mg Lactic acid 50 mg Sodium hydroxide q.s. forpH 4.5 Water q.s. 100 g

TABLE 10 10 mM gel compound B 329.40 mg HEC 2 g Glycerol 5 g Methylparaben 180 mg Propyl paraben 20 mg Lactic acid 50 mg Sodium hydroxideq.s. for pH 4.5 Water q.s. 100 g

Various modifications and alterations to the present invention may beappreciated based on a review of this disclosure, These changes andadditions are intended to be within the scope and spirit of thisinvention as defined by the following claims.

1-27. (canceled)
 28. A method for treating chancroids in a patient inneed thereof, comprising administering to the patient a compound thatis:4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4-dichlorophenyl)methyl]-6-[(4-hydroxybutyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(3-hydroxypropyl)amino]-2-pyrimidinyl]amino]-benzonitrile;N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]acetamide;N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]butanamide;4-[[2-amino-6-(2,6-dichlorophenoxy)-4-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2-hydroxy-2-phenylethyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[3-(2-oxo-1-pyrrolidinyl)propyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(2-hydroxyethoxy)ethyl]amino]-2-pyrimidinyl]amino]benzonitrilemonohydrochloride;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2,3-dihydroxypropyl)amino]-2-pyrimidinyl]-amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-(hydroxyamino)-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2-cyanoethyl)amino]-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(1-pyrrolidinyl)ethyl]amino]-2-pyrimidinyl]-amino]benzonitrile;4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-(2,4,6-trimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dichloro-4-(trifluoromethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4-dichloro-6-methylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(cyanophenyl)amino]-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;4-[[4-[(2,4-dibromo-6-fluorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[methyl(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)amino-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-6-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-amino-6-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-(2-bromo-4-chloro-6-methylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4[(4-chloro-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;3,5-dichloro-4-[[2-[(4-cyanophenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dichloro-4-(trifluoromethoxy)phenyl]amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[4-[(2,4-dibromo-3,6-dichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dibromo-4-propylphenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(4-(1,1-dimethylethyl)-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[2-[(4-cyanophenyl)amino]-4-pyrimidinyl]oxy]-3,5-dimethylbenzonitrile;4-[[4-[(4-chloro-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino-3,5-dimethylbenzonitrile;4-[[4-[[4-(1,1-dimethylethyl)-2,6-dimethylphenyl]amino]-5-methyl-2-pyrimidinyl]-amino]benzonitrile;4-[[4-[(4-bromo-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[5-methyl-4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dibromo-4-propylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;4-[[4-[(phenylmethyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[5-chloro-4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[5-bromo-4-(4-cyano-2,6-dimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[5-bromo-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile;or4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile;or an N-oxide, a pharmaceutically acceptable addition salt, astereochemically isomeric form thereof, or a quaternary amine thereof.29. The method of claim 28, wherein the compound is:4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile,or4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 30. The method of claim 29,wherein the compound is4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile,or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 31. The method of claim 30,wherein the compound is4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile.32. The method of claim 29, wherein the compound is4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile,or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 33. The method of claim 31,wherein the compound is4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile.34. The method of claim 28, wherein the compound is administered to thevagina.
 35. A method for treating chancroids in a patient in needthereof, comprising administering to the patient a compositioncomprising a compound that is:4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4-dichlorophenyl)methyl]-6-[(4-hydroxybutyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(3-hydroxypropyl)amino]-2-pyrimidinyl]amino]-benzonitrile;N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]acetamide;N-[2-[(4-cyanophenyl)amino]-6-[(2,6-dichlorophenyl)methyl]-4-pyrimidinyl]butanamide;4-[[2-amino-6-(2,6-dichlorophenoxy)-4-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2-hydroxy-2-phenylethyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[3-(2-oxo-1-pyrrolidinyl)propyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(2-hydroxyethoxy)ethyl]amino]-2-pyrimidinyl]amino]benzonitrilemonohydrochloride;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[(2,3-dihydroxypropyl)amino]-2-pyrimidinyl]-amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-(hydroxyamino)-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2-cyanoethyl)amino]-6-[(2,6-dichlorophenyl)methyl]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)methyl]-6-[[2-(1-pyrrolidinyl)ethyl]amino]-2-pyrimidinyl]-amino]benzonitrile;4-[[4-amino-6-[(2,6-dichlorophenyl)methyl]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-(2,4,6-trimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dichlorophenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dichloro-4-(trifluoromethyl)phenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4-dichloro-6-methylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(cyanophenyl)amino]-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;4-[[4-[(2,4-dibromo-6-fluorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[methyl(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,4,6-trimethylphenyl)amino-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-6-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[2-amino-6-[(2,4,6-trimethylphenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-(2-bromo-4-chloro-6-methylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(4-chloro-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;3,5-dichloro-4-[[2-[(4-cyanophenyl)amino]-4-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dichloro-4-(trifluoromethoxy)phenyl]amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[4-[(2,4-dibromo-3,6-dichlorophenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dibromo-4-propylphenyl]amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(4-(1,1-dimethylethyl)-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[2-[(4-cyanophenyl)amino]-4-pyrimidinyl]oxy]-3,5-dimethylbenzonitrile;4-[[4-[(4-chloro-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino-3,5-dimethylbenzonitrile;4-[[4-[[4-(1,1-dimethylethyl)-2,6-dimethylphenyl]amino]-5-methyl-2-pyrimidinyl]-amino]benzonitrile;4-[[4-[(4-bromo-2,6-dimethylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[5-methyl-4-[(2,4,6-trimethylphenyl)thio]-2-pyrimidinyl]amino]benzonitrile;4-[[4-[(2,6-dibromo-4-propylphenyl)amino]-5-methyl-2-pyrimidinyl]amino]benzonitrile;4-[[4-[[2,6-dibromo-4-(1-methylethyl)phenyl]amino]-5-methyl-2-pyrimidinyl]amino]-benzonitrile;4-[[2-[(4-cyanophenyl)amino]-5-methyl-4-pyrimidinyl]amino]-3,5-dimethylbenzonitrile;4[[4-[(phenylmethyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[5-chloro-4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[5-bromo-4-(4-cyano-2,6-dimethylphenoxy)-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile;4-[[5-bromo-6-[(4-cyano-2,6-dimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;4-[[4-amino-5-chloro-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile;or4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]benzonitrile; or an N-oxide, a pharmaceutically acceptable additionsalt, a stereochemically isomeric form thereof, or a quaternary aminethereof.
 36. The method of claim 35, wherein the compound is:4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile,or4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 37. The method of claim 36,wherein the compound is4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile;or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 38. The method of claim 37,wherein the compound is4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]benzonitrile.39. The method of claim 36, wherein the compound is4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile,or an N-oxide thereof, a pharmaceutically acceptable addition saltthereof, or a quaternary amine thereof.
 40. The method of claim 39,wherein the compound is4-[[4-amino-5-bromo-6-(4-cyano-2,6-dimethylphenyloxy)-2-pyrimidinyl]amino]-benzonitrile.41. The method of claim 35, wherein the composition is administered tothe vagina.
 42. The method of claim 35, wherein the composition isadministered in the form of an intravaginal ring.
 43. The method ofclaim 35, wherein the composition is administered in a topical form. 44.The method of claim 35, wherein the composition is administered in theform of a bioadhesive.
 45. The method of claim 35, wherein thecomposition is administered is in the form of an immediate release drugdelivery system.
 46. The method of claim 35, wherein the composition isadministered in the form of a sustained release drug delivery system.47. The method of claim 35, wherein the composition is a gel and furthercomprises: a gel-forming compound, a buffer, a pharmaceuticallyacceptable diluent, optionally a humectant, and optionally apreservative.
 48. The method of claim 35, wherein the compositioncomprises4-[[4-[(2,4,6-trimethylphenyl)amino]-2-pyrimidinyl]amino]-benzonitrile,hydroxyethyl cellulose, glycerol, methyl paraben, propyl paraben, lacticacid, sodium hydroxide, and water.
 49. The method of claim 35, whereinthe composition comprises one or more additional antiretroviralcompounds.
 50. The method of claim 35, wherein the composition comprisesone or more components selected from an antibody, a detergent orsurfactant, a coating for the pathogen, a coating for the site oftransmission, an antibiotic peptide or a pH regulator.
 51. The method ofclaim 35, wherein the composition comprises a spermicidal compound.